Curcumin analogs with anti-tumor and anti-angiogenic properties

ABSTRACT

The present invention is directed to curcumin analogs exhibiting anti-tumor and anti-angiogenic properties, pharmaceutical formulations including such compounds and methods of using such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional No.60/168,913, filed Dec. 3, 1999, which is incorporated by referenceherein in its entirety.

FIELD OF THE INVENTION

[0002] The invention relates to compounds useful for the treatment ofcancer, and in particular to compounds exhibiting anti-tumor andanti-angiogenic properties and methods for using such compounds.

BACKGROUND OF THE INVENTION

[0003] Tissue factor (TF) is a sedimentable, integral membrane receptorprotein with an estimated molecular weight of 42-47 kDa. Peritumorfibrin deposition, which is characteristic of most types of humancancer, is the result of the local expression of potent procoagulantslike tissue factor (TF) in tumor cells, tumor-associated macrophages(TAMs) and tumor-associated vascular endothelial cells (VECs). Inaddition to the importance of TF expression in the pathogenesis of thethrombotic complications common to cancer patients, increasing evidencelinks TF expression to the regulation of tumor angiogenesis, growth andmetastasis. For example, angiogenesis in vivo is inhibited by TFantisense. Further, murine tumor cells transfected to overexpress TFenhance vascular permeability factor (VEGF) transcription andtranslation. Conversely, tumor cells transfected with TF antisensereduce VEGF transcription and translation. VEGF acts specifically onVECs to promote vascular permeability, endothelial cell growth andangiogenesis, and has been shown to induce expression of TF activity inVECs and monocytes and is chemotactic for monocytes, osteoblasts andVECs. Expression of TF and VEGF in cancer cells is further enhancedunder hypoxic condition. Thus, there is evidence to suggest that TF is akey molecule participating in the regulation of VEGF synthesis and,hence, tumor angiogenesis in cancer.

[0004] Relatively few compounds exhibiting anti-angiogenic propertiesuseful in the treatment of cancer have been investigated. Curcumin(diferuloylmethane), the aromatic yellow pigment in curry, turmeric andmustard, is known to have anti-angiogenic, anti-tumor, and anti-tumorpromoting properties. In addition, curcumin exhibits numerous othertherapeutic effects, including anti-oxidative, anti-thrombotic,anti-inflammatory, anti-cholesterol and anti-diabetic properties. Twoother compounds that have received considerable attention are genistein,a soybean-derived isoflavone tyrosine kinase C inhibitor, and linomide,a quinoline-3-carboxaminde. Certain flavonoids, such as apigenin, havebeen shown to be more potent inhibitors of cell proliferation and invitro angiogenesis than genistein. There remains a need in the art forcompounds that exhibit anti-tumor and anti-angiogenic properties for usein cancer therapy.

SUMMARY OF THE INVENTION

[0005] The present invention provides a group of curcumin analogs thatinhibit TF expression and VEGF expression in cancer cells and invascular endothelial cells in the tumor microenvironment, therebyblocking tumor angiogenesis and growth, without exhibiting a high levelof toxicity with regard to normal vascular endothelial cells. Theanti-angiogenic and anti-tumor compounds of the present invention can beuseful in the treatment of any condition benefiting from angiogenesisinhibition, such as cancer.

[0006] In one aspect, the present invention provides compounds ofFormula (I) below.

[0007] In another aspect, the present invention provides apharmaceutical formulation comprising a compound of Formula (I) orFormula (II) below in a pharmaceutically acceptable carrier.

[0008] In a third aspect, the present invention provides a method oftreating cancerous tissue in a subject, comprising administering aneffective amount of a compound of Formula (I) or Formula (II) to thesubject. Preferably, the compound is administered in a pharmaceuticallyacceptable carrier. The effective amount is preferably an amountsufficient to inhibit VEGF or TF production in the cancerous tissue.

BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Having thus described the invention in general terms, referencewill now be made to the accompanying drawings, wherein:

[0010]FIGS. 1A and 1B graphically illustrate the relationship betweencell viability and VEGF production of human melanoma cells aftertreatment with the compounds of the present invention at twoconcentrations;

[0011]FIGS. 2A and 2B graphically illustrate the effect of knowncompounds and the compounds of the present invention on human melanomacell viability;

[0012]FIGS. 3A and 3B graphically illustrate the effect of knowncompounds and the compounds of the present invention on human breastcancer cell viability;

[0013]FIGS. 4A and 4B graphically illustrate the effect of knowncompounds and the compounds of the present invention on transformedmurine endothelial cell viability; and

[0014]FIGS. 5A and 5B graphically illustrate the effect of knowncompounds and the compounds of the present invention on humanendothelial cell viability.

DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention will be described more fully hereinafter,including preferred embodiments thereof. This invention may, however, beembodied in many different forms and should not be construed as limitedto the embodiments set forth herein; rather, these embodiments areprovided so that this disclosure will be thorough and complete, and willfully convey the scope of the invention to those skilled in the art.

[0016] As used herein, the term “compound” is intended to refer to achemical entity, whether in the solid, liquid or gaseous phase, andwhether in a crude mixture or purified and isolated. The terms “alkyl,”“alkene,” and “alkoxy” include straight chain and branched alkyl,alkene, and alkoxy, respectively. The term “lower alkyl” refers to C1-C4alkyl. The term “alkoxy” refers to oxygen substituted alkyl, forexample, of the formulas —OR or —ROR¹, wherein R and R¹ are eachindependently selected alkyl. The terms “substituted alkyl” and“substituted alkene” refer to alkyl and alkene, respectively,substituted with one or more non-interfering substituents, such as butnot limited to, C3-C6 cycloalkyl, e.g., cyclopropyl, cyclobutyl, and thelike; acetylene; cyano; alkoxy, e.g., methoxy, ethoxy, and the like;lower alkanoyloxy, e.g., acetoxy; hydroxy; carboxyl; amino; loweralkylamino, e.g., methylamino; ketone; halo, e.g. chloro or bromo;phenyl; substituted phenyl, and the like. The term “halogen” includesfluorine, chlorine, iodine and bromine.

[0017] “Aryl” means one or more aromatic rings, each of 5 or 6 carbonatoms. Multiple aryl rings may be fused, as in naphthyl or unfused, asin biphenyl. Aryl rings may also be fused or unfused with one or morecyclic hydrocarbon, heteroaryl, or heterocyclic rings.

[0018] “Substituted aryl” is aryl having one or more non-interferinggroups as substituents.

[0019] “Heteroaryl” is an aryl group containing from one to four N, O,or S atoms(s) or a combination thereof, which heteroaryl group isoptionally substituted at carbon or nitrogen atom(s) with C1-6 alkyl,—CF₃, phenyl, benzyl, or thienyl, or a carbon atom in the heteroarylgroup together with an oxygen atom form a carbonyl group, or whichheteroaryl group is optionally fused with a phenyl ring. Heteroarylrings may also be fused with one or more cyclic hydrocarbon,heterocyclic, aryl, or heteroaryl rings. Heteroaryl includes, but is notlimited to, 5-membered heteroaryls having one hetero atom (e.g.,thiophenes, pyrroles, furans); 5 membered heteroaryls having twoheteroatoms in 1,2 or 1,3 positions (e.g., oxazoles, pyrazoles,imidazoles, thiazoles, purines); 5-membered heteroaryls having threeheteroatoms (e.g., triazoles, thiadiazoles); 5-membered heteroarylshaving 3 heteroatoms; 6-membered heteroaryls with one heteroatom (e.g.,pyridine, quinoline, isoquinoline, phenanthrine,5,6-cycloheptenopyridine); 6-membered heteroaryls with two heteroatoms(e.g., pyridazines, cinnolines, phthalazines, pyrazines, pyrimidines,quinazolines); 6-membered heretoaryls with three heteroatoms (e.g.,1,3,5-triazine); and 6-membered heteroaryls with four heteroatoms.

[0020] “Substituted heteroaryl” is heteroaryl having one or morenon-interfering groups as substituents.

[0021] “Heterocycle” or “heterocyclic” means one or more rings of 5, 6or 7 atoms with or without unsaturation or aromatic character and atleast one ring atom which is not carbon. Preferred heteroatoms includesulfur, oxygen, and nitrogen. Multiple rings may be fused, as inquinoline or benzofuran.

[0022] “Substituted heterocycle” is heterocycle having one or more sidechains formed from non-interfering substituents.

[0023] “Non-interfering substituents” are those groups that yield stablecompounds. Suitable non-interfering substituents or radicals include,but are not limited to, halo, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, C₁-C₁₀ alkoxy, C₇-C₁₂ aralkyl, C₇-C₁₂ alkaryl, C₃-C₁₀cycloalkyl, C₃-C₁₀ cycloalkenyl, phenyl, substituted phenyl, toluoyl,xylenyl, biphenyl, C₂-₁₂ alkoxyalkyl, C₇-C₁₂ alkoxyaryl, C₇-C₁₂aryloxyalkyl, C₆-C₁₂ oxyaryl, C₁-C₆ alkylsulfinyl, C₁-C₁₀ alkylsulfonyl,—(CH₂)_(m)—O—(C₁-C₁₀ alkyl) wherein m is from 1 to 8, aryl, substitutedaryl, substituted alkoxy, fluoroalkyl, heterocyclic radical, substitutedheterocyclic radical, nitroalkyl, —NO₂, —CN, —NRC(O)—(C₁-C₁₀ alkyl),—C(O)—(C₁-C₁₀ alkyl), C₂-C₁₀ thioalkyl, —C(O)O—(C₁-C₁₀ alkyl), —OH,—SO₂, ═S, —COOH, —NR₂, carbonyl, —C(O)—(C₁-C₁₀ alkyl)—CF₃, —C(O)—CF₃,—C(O)NR₂, —(C₁-C₁₀ alkyl)—S—(C₆-C₁₂ aryl), —C(O)—(C₆—C₁₂ aryl),—(CH₂)_(m)—O—(CH₂)_(m)—O—(C₁-C₁₀ alkyl) wherein each m is from 1 to 8,—C(O)NR₂, —C(S)NR₂, —SO₂NR₂, —NRC(O)NR₂, —NRC(S)NR₂, salts thereof, andthe like. Each R as used herein is H, alkyl or substituted alkyl, arylor substituted aryl, aralkyl, or alkaryl.

[0024] The present invention provides compounds of Formula (I)

[0025] wherein:

[0026] Y is OH, halogen, or CF₃;

[0027] Z is H, OH, OR₁, halogen, or CF₃;

[0028] X₁ and X₂ are independently C or N; and

[0029] A is selected from the group consisting of:

[0030] wherein n is 1-8; X₃ is O, S, SO, SO₂, NH, or NR₁; Q is NH orNR₁; and V₁₋₄ are each independently OH, OR₂, or halogen; R₁ and R₂ areindependently H, alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocycle, substitutedheterocycle, acyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl ordialkylaminocarbonyl; the dashed lines indicate the presence of optionaldouble bonds; and L is the point of bonding of A to the compoundstructure, with the proviso that Z is not H when Y is OH, Cl or Br and Ais

[0031] and pharmaceutically acceptable salts thereof.

[0032] The present invention also provides a method of treatingcancerous tissue in a subject, such as a human or other mammal,comprising administering to the subject an effective amount of acompound of Formula (I) above or a compound of Formula (II)

[0033] wherein:

[0034] X₄ is (CH₂)_(m), O, S, SO, SO₂, or NR₁₂, where R₁₂ is H, alkyl,substituted alkyl, acyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl or dialkylaminocarbonyl;

[0035] m is 1-7;

[0036] each X₅ is independently N or C—R₁₁;

[0037] and each R₃-R₁₁ are independently H, halogen, hydroxyl, alkoxy,CF₃, alkyl, substituted alkyl, alkenyl, alkynyl, cycloalkyl, substitutedcycloalkyl, aryl, substituted aryl, alkaryl, arylalkyl, heteroaryl,substituted heteroaryl, heterocycle, substituted heterocycle, amino,alkylamino, dialkylamino, carboxylic acid, carboxylic ester,carboxamide, nitro, cyano, azide, alkylcarbonyl, acyl, ortrialkylammonium; and

[0038] the dashed lines indicate optional double bonds;

[0039] with the proviso that when X₄ is (CH₂)_(m), m is 2-6, and each X₅is C—R₁₁, R₃-R₁₁ are not alkoxy, and when X₄ is NR₁₂ and each X₅ is N,R₃-R₁₀ are not alkoxy, alkyl, substituted alkyl, alkenyl, alkynyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, alkaryl,arylalkyl, heteroaryl, substituted heteroaryl, amino, alkylamino,dialkylamino, carboxylic acid, or alkylcarbonyl.

[0040] The present invention includes all stereoisomeric configurationsof the compounds of Formula (I) and Formula (II), including both opticalisomers, such as enantiomers and diastereoisomers, and geometric(cis-trans) isomers.

[0041] Examples of the compounds of the present invention include, butare not limited to:

(EF2) R = 2-OH (EF4)  X = C, R = 2-OH (EF3) R = 3-OH (EF31) X = C-1, R =2-OH (EF1) R = 4-OH (EF25) X = O, R = 2-OH (EF8) R = 2-F (EF29) X = O, R= 2-F (EF9) R = 2,4-F (EF30) X = O, R = 2,4-F (EF10) R = 3,4-F (EF36) X= O, R = 3,4(OMe) (EF23) R = 2,6-F (EF28) X = O, R = 2-OMe (MD6) R =3,4-(OMe) (EF27) X = O, R = 4-OMe (EF16) R = 2-OMe (EF34) X = NMe, R =2-OH (EF17) R = 3-OMe (EF33) X = NMe, R = 2-F (EF18) R = 4-OMe (EF47) X= NMe, R = 2,4-F (EF35) X = NMe, R = 3,4-(OMe) (EF24) X = NH₂OAc, R =2-F (EF26) X = NH₂Cl, R = H

(EF15)n = 0, R = H (EF32) R = 2-OH (EF13)n = 1, R = H, (E, E) (EF48) R =2-F (EF14)n = 1, R = H, (E, Z) (EF19) R = 2,4-F₂ (EF11)n = 2, R = 2-OH(EF20) R = 3,4-F₂ (EF12)n = 2, R = H (MD279L) R = 3,4-(OMe)

(EF37) R₁ = Me, R = 3,4-F (EF21) R = 2,4-F (MD38) R₁ = Me, R = 3,4-(OMe)(EF22) R = 3,4-F (EF44) R₁ = Pr, R = 3,4-(OMe) (MD279U) R = 3,4-(OMe)

(EF46) R₁ = CH₂Br, R = 3,4-(OMe) (EF40) R₁ = Et, R = 3,4-F (EF41) R₁ =Et, R = 3,4-(OMe) (EF49) R₁ = Ph, R = 2-F (EF39) (EF45) (EF43) R₁ = Ph,R₁ = morph. R₁ = menth., R 3,4-(OMe) R = 3,4-(OMe) R 3,4-(OMe)

(EF42) R = 3,4-(OMe) X = Cl Y = H (EF50) R = 3,4-(OMe) X = OH Y = Me

[0042]

[0043] and the like and pharmaceutically acceptable salts thereof.Additional exemplary compounds are given in the appended examples.

[0044] The compounds of the present invention may be prepared accordingto methods known in the art, particularly in light of the disclosure andexamples set forth herein. The starting materials used to synthesize thecompounds of the present invention are commercially available or capableof preparation using methods known in the art. For example, somecompounds of the present invention may be prepared by reaction of anaromatic aldehyde, such as hydroxybenzaldehyde or fluoro-substitutedbenzaldehyde, with a ketone, such as acetone, cyclohexanone,cyclopentanone, tetrahydro-4-H-pyran-4-one, N-methyl-4-piperidone,piperidin-4-one, and the like, under basic aldol condensationconditions. Similarly, other compounds of the present invention may beprepared by reaction of an alkoxy-substituted benzaldehyde oranisaldehyde with a ketone. As would be understood, the actual ketone oraldehyde utilized will depend on the type and position of thesubstituents of the desired final compound. The salts of the presentinvention may be prepared, in general, by reaction of a compound of theinvention with the desired acid or base in solution. After the reactionis complete, the salts can be crystallized from solution by the additionof an appropriate amount of solvent in which the salt is insoluble.

[0045] The compounds of Formula (I) or Formula (II) can havepharmaceutical activity and can be useful in the treatment of a subjectsuffering from one or more conditions that would benefit from inhibitionof angiogenesis. For example, the compounds of the present invention canbe used in the treatment of cancerous tissue and the tumors associatedtherewith, including breast, colon, prostate and skin cancer. Inaddition, the compounds of the present invention can be useful formediating inflammation, rheumatoid arthritis and certain forms ofdiabetes. Subjects which can be treated include animal subjects,typically vertebrates, including both mammalian (e.g., human, cat, dog,cow, horse, sheep, pig, monkey, ape, etc.) and avian subjects (e.g.,chicken, turkey, duck, goose, quail, pheasant, etc.). It is believed,for example, that administering an effective amount of a compound ofFormula (I) or Formula (II) to a subject can result in inhibition ofangiogenesis in cancerous tissue. Thus, the present invention canprovide methods for treating tumor-bearing subjects in which thecompounds of the invention are administered to the subject in need ofsuch treatment in an amount effective and in a manner effective tocombat such tumors, for example, by virtue of inhibition of angiogenesiswithin the tumor. The anti-angiogenesis effect is believed to result, atleast in part, from inhibition of TF and/or VEGF production in thetumor. In addition, it is believed that the compounds of the presentinvention can be used as a prophylactic treatment to prevent certaintypes of inflammatory skin conditions including, but not limited to,dermatitis and mild cases of skin cancer.

[0046] The compounds of Formula (I) or Formula (II) may be administeredper se or in the form of a pharmaceutically acceptable salt. When usedin medicine, the salts of the compounds of Formula (I) or Formula (II)should be both pharmacologically and pharmaceutically acceptable, butnon-pharmaceutically acceptable salts may conveniently be used toprepare the free active compound or pharmaceutically acceptable saltsthereof and are not excluded from the scope of this invention. Suchpharmacologically and pharmaceutically acceptable salts can be preparedby reaction of a compound of Formula (I) or Formula (II) with an organicor inorganic acid, using standard methods detailed in the literature.Examples of useful salts include, but are not limited to, those preparedfrom the following acids: hydrochloric, hydrobromic, sulfuric, nitric,phosphoric, maleic, acetic, salicyclic, p-toluenesulfonic, tartaric,citric, methanesulphonic, formic, malonic, succinic,naphthalene-2-sulphonic and benzenesulphonic, and the like. Also,pharmaceutically acceptable salts can be prepared as alkaline metal oralkaline earth salts, such as sodium, potassium, or calcium salts of thecarboxylic acid group.

[0047] Thus the present invention also provides pharmaceuticalformulations or compositions, both for veterinary and for human medicaluse, which comprise the compounds of Formula (I) or Formula (II) or apharmaceutically acceptable salt thereof with one or morepharmaceutically acceptable carriers thereof and optionally any othertherapeutic ingredients, such as other chemotherapeutic agents. Thecarrier(s) must be pharmaceutically acceptable in the sense of beingcompatible with the other ingredients of the formulation and not undulydeleterious to the recipient thereof.

[0048] The compositions includes those suitable for oral, rectal,topical, nasal, ophthalmic, or parenteral (including intraperitoneal,intravenous, subcutaneous, or intramuscular injection) administration.The compositions may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active agent intoassociation with a carrier that constitutes one or more accessoryingredients. In general, the compositions are prepared by uniformly andintimately bringing the active compound into association with a liquidcarrier, a finely divided solid carrier or both, and then, if necessary,shaping the product into desired formulations.

[0049] Compositions of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets, tablets, lozenges, and the like, each containing apredetermined amount of the active agent as a powder or granules; or asuspension in an aqueous liquor or non-aqueous liquid such as a syrup,an elixir, an emulsion, a draught, and the like.

[0050] A tablet may be made by compression or molding, optionally withone or more accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine, with the active compound being in afree-flowing form such as a powder or granules which is optionally mixedwith a binder, disintegrant, lubricant, inert diluent, surface activeagent or dispersing agent. Molded tablets comprised with a suitablecarrier may be made by molding in a suitable machine.

[0051] A syrup may be made by adding the active compound to aconcentrated aqueous solution of a sugar, for example sucrose, to whichmay also be added any accessory ingredient(s). Such accessoryingredients may include flavorings, suitable preservatives, an agent toretard crystallization of the sugar, and an agent to increase thesolubility of any other ingredient, such as polyhydric alcohol, forexample, glycerol or sorbitol.

[0052] Formulations suitable for parental administration convenientlycomprise a sterile aqueous preparation of the active compound, which canbe isotonic with the blood of the recipient.

[0053] Nasal spray formulations comprise purified aqueous solutions ofthe active agent with preservative agents and isotonic agents. Suchformulations are preferably adjusted to a pH and isotonic statecompatible with the nasal mucous membranes.

[0054] Formulations for rectal administration may be presented as asuppository with a suitable carrier such as cocoa butter, orhydrogenated fats or hydrogenated fatty carboxylic acids.

[0055] Ophthalmic formulations are prepared by a similar method to thenasal spray, except that the pH and isotonic factors are preferablyadjusted to match that of the eye.

[0056] Topical formulations comprise the active compound dissolved orsuspended in one or more media such as mineral oil, petroleum,polyhydroxy alcohols or other bases used for topical formulations. Theaddition of other accessory ingredients as noted above may be desirable.

[0057] Further, the present invention provides liposomal formulations ofthe compounds of Formula (I) or Formula (II) and salts thereof. Thetechnology for forming liposomal suspensions is well known in the art.When the compound of Formula (I) or Formula (II) or salt thereof is anaqueous-soluble salt, using conventional liposome technology, the samemay be incorporated into lipid vesicles. In such an instance, due to thewater solubility of the compound or salt, the compound or salt will besubstantially entrained within the hydrophilic center or core of theliposomes. The lipid layer employed may be of any conventionalcomposition and may either contain cholesterol or may becholesterol-free. When the compound or salt of interest iswater-insoluble, again employing conventional liposome formationtechnology, the salt may be substantially entrained within thehydrophobic lipid bilayer that forms the structure of the liposome. Ineither instance, the liposomes that are produced may be reduced in size,as through the use of standard sonication and homogenization techniques.The liposomal formulations containing the compounds of Formula (I) orFormula (II) or salts thereof, may be lyophilized to produce alyophilizate which may be reconstituted with a pharmaceuticallyacceptable carrier, such as water, to regenerate a liposomal suspension.

[0058] Pharmaceutical formulations are also provided which are suitablefor administration as an aerosol, by inhalation. These formulationscomprise a solution or suspension of the desired compound of Formula (I)or Formula (II) or a salt thereof or a plurality of solid particles ofthe compound or salt. The desired formulation may be placed in a smallchamber and nebulized. Nebulization may be accomplished by compressedair or by ultrasonic energy to form a plurality of liquid droplets orsolid particles comprising the compounds or salts.

[0059] In addition to the aforementioned ingredients, the compositionsof the invention may further include one or more accessory ingredient(s)selected from the group consisting of diluents, buffers, flavoringagents, binders, disintegrants, surface active agents, thickeners,lubricants, preservatives (including antioxidants) and the like.

[0060] Preferably, for purposes of cancer therapy, a compound of Formula(I) or Formula (II) is administered to the subject in an amountsufficient to inhibit production of TF or VEGF, thereby inhibitingangiogenesis. However, the therapeutically effective dosage of anyspecific compound will vary somewhat from compound to compound, patientto patient, and will depend upon the condition of the patient and theroute of delivery. As a general proposition, a dosage from about 0.5 toabout 20 mg/kg body weight, preferably from about 1.0 to about 5.0mg/kg, will have therapeutic efficacy. When administered conjointly withother pharmaceutically active agents, even less of the compounds ofFormula (I) or Formula (II) may be therapeutically effective. Thecompound of Formula (I) or Formula (II) may be administered once orseveral times a day. The duration of the treatment may be once per dayfor a period of from two to three weeks and may continue for a period ofmonths or even years. The daily dose can be administered either by asingle dose in the form of an individual dosage unit or several smallerdosage units or by multiple administration of subdivided dosages atcertain intervals.

[0061] For the following examples, RPMI-7951 human melanoma, MDA-MB-231and MDA-MB-435 human breast cancer cell lines were purchased fromAmerican Type Cell Collection (Rockville, Md.). HUVECs were obtainedfrom the Department of Dermatology, Emory University. Murine endothelialcells infected with simian virus 40 (SV40) large T antigen and activatedH-ras (SVR), were a kind gift from Dr. Jack Arbiser at Emory. RPMI-7951,MDA-MB-231 and MDA-MB-435 cell lines were cultured in MEM-alpha medium(GIBCO-BRL, Long Island, N.Y.) containing 10% fetal bovine serum(RPMI-7951, MDA-MB-231) or 5% FBS (MDA-MB-435) at 37° C. and under 5%CO₂/95% air. SVR cells were cultured in DMEM (Mediatech cellgro)containing 10% FBS and 2mM L-glutamine. Complete HUVEC media was a giftfrom the Cell Culture Center in the Department of Dermatology, EmoryUniversity. The cells were cultured in 48 well plates in all of theexperiments described.

[0062] Neutral Red Assay was utilized to determine the effect of thecompounds of the present invention on cell viability. Neutral Red waspurchased from GIBCO-BRL (Long Island, N.Y.). Cells were plated at aconcentration of 20,000 cells/well and cultured overnight. Compounds orvehicle (DMSO 0.1%) were then added and the plates were incubated for 72hours. Supernatant from each well was either aspirated or collected andmedia containing Neutral Red (GIBCO-BRL, Long Island, N.Y.) at aconcentration of 15 μl/ml was then added to each well. The plates werethen incubated at 37° C. for 30 minutes. Next, the cells were washedwith twice with PBS and alcoholic-HCl (0.5N-HCl/35% ethanol) was addedto each well. The plates were then placed on a plate shaker until allresidues were solubilized (pink color). The solubilized mixtures werethen transferred to a 96 well plate and the absorbances were read on amicro-test plate reader at a wavelength of 570 nM.

[0063] A VEGF enzyme-linked immunosorbent assay (ELISA) was utilized todetermine the effect of the compounds of the present invention on VEGFproduction of a variety of human cancer cell lines. For the VEGF assay,cells were plated at a concentration of 80,000 cells/well and culturedovernight. Compounds or vehicle (DMSO 0.1%) were then added and theplates were incubated for 72 hours. Supernatant was then collected fromeach well and frozen in a −80° C. freezer until needed. Cell viabilitywas determined by Neutral Red Assay. VEGF ELISA Kit (R & D, Minneapolis,Minn.) was used to determine the amount of VEGF in the culturesupernatants. The ELISA was carried out according to the manufacturer'sprocedure.

[0064] A TF ELISA assay was utilized to determine the effect of thecompounds of the present invention on TF production of human cancer celllines. For the TF assay, cells were plated at a concentration of 80,000cells/well and cultured overnight. Compounds or vehicle (DMSO 0.1%) werethen added and the plates were incubated for 72 hours. Cells weretreated with 1% Triton X-100 in PBS and left overnight at 4° C.overnight to solubilize TF. Supernatant was then collected from eachwell and frozen until needed. IMUBIND Tissue Factor ELISA Kit (AmericanDiagnostica Inc, Greenwich, Conn.) was used to determine TFconcentration in each sample. The ELISA was carried out according tomanufacturer's procedure.

EXPERIMENTAL Example 1 Preparation of EF1, EF2, EF3, EF4, EF25, EF31,EF34 compounds

[0065] The compounds of this series were all synthesized by thefollowing procedure: Aqueous NaOH (20 wt %, 15 ml, 75 mmol) was addeddropwise to a vigorously stirred solution of hydroxybenzaldehyde (51mmol) and ketone (25 mmol) in EtOH abs (20 mL). The reaction was stirredat room temperature for 48 hrs, H₂O dist (100 mL) was added, and thepurple solution was neutralized by gently bubbling CO₂ through it. Theprecipitating yellow solid was filtered off, washed with H₂O dist anddried under vacuum. The products were purified by recrystallization. Adescription of each compound obtained by the above process is givenbelow.

[0066] 1,5-Bis(4-hydroxyphenyl)penta-1,4-dien-3-one (EF1): yellow solid(6%), mp 236° C. (acetone/H₂O).

[0067]¹H NMR (400 MHz, CD₃OD) δ 7.71 (2H, d, J=16 Hz), 7.58 (4H, d,J=8.8 Hz), 7.07 (2H, d, J=16 Hz), 6.84 (4H, d, J=8.4 Hz). ¹³C NMR (100MHz, CD₃OD) δ 191.9, 161.8, 145.3, 131.8, 127.8, 123.6, 117.1. EIHRMS:m/z266.0943 (M⁺, C₁₇H₁₄O₃ requires 266.0943).

[0068] 1,5-Bis(2-hydroxyphenyl)penta-1,4-dien-3-one (EF2): yellow solid(75%), mp 155° C. (acetone/H₂O).

[0069]¹H NMR (400 MHz, CD₃OD) δ 8.09 (2H, d, J=16 Hz,), 7.63 (2H, dd,J=8.4 Hz, J=1.6 Hz), 7.31 (2H, d, J=16 Hz), 7.24 (2H, td, J=7.6 Hz,J=1.6 Hz), 6.88 (t, 4H, J=7.2 Hz). ¹³C NMR (100 MHz, CD₃OD) δ 192.8,158.9, 141.0, 133.1, 130.2, 126.3, 123.2, 121.0, 117.2. Anal. Calcd forC₁₇H₁₄O₃: C, 78.68; H, 5.30. Found: C, 76.56; H, 5.32. EIHRMS: m/z248.0837 ((M—H₂O)⁺, C₁₇H₁₂O₂ requires 248.0837).

[0070] 1,5-Bis(3-hydroxyphenyl)penta-1,4-dien-3-one (EF3): yellow solid(15%), mp 198-200° C. (acetone/H₂O).

[0071]¹H NMR (400 MHz, CD₃OD) δ 7.70 (2H, d, J=16 Hz), 7.24 (2H, t,J=7.6 Hz), 7.17 (2H, d, J=16 Hz), 7.17 (2H, d, J=8 Hz), 7.11 (2H, s),6.73 (2H, dd, J=8 Hz, J=2.4 Hz). ¹³C NMR (100 MHz, CD₃OD) δ 191.6,159.3, 145.5, 137.6, 131.2, 126.4, 121.4, 119.1, 115.9. Anal. Calcd forC₁₇H₁₄O₃: C, 78.68; H, 5.30. Found: C, 76.41: H, 5.48. EIHRMS: m/z266.0943 (M⁺, C₁₇H₁₄O₃ requires 266.0943).

[0072] 2,6-Bis(2-hydroxybenzylidene)cyclohexanone (EF4): yellow solid(70%, recrystallized from acetone/H₂O).

[0073]¹H NMR (400 MHz, CD₃OD) δ 7.98 (2H, s,), 7.32 (2H, dd, J=7.6 Hz,J=1.2 Hz), 7.19 (2H, td, J=7.6 Hz, J=1.2 Hz), 6.86 (4H, m), 2.86 (4H,m), 1.75 (2H, m). ¹³C (100 MHz, CD₃OD) δ 192.8, 158.2, 137.3, 134.5,131.5, 124.5, 120.2, 116.6, 29.9, 24.8. Anal. Calcd for C₂₀H₁₈O₃: C,78.41; H, 5.92. Found: C, 78.15; H, 6.03. EIHRMS: m/z 306.1263 (M⁺,C₂₀H₁₈O₃ requires 306.1256).

[0074] 3,5-Bis(2-hydroxybenzylidene)tetrahydro-4-H-pyran-4-one (EF25):yellow solid (60%, recrystallized from acetone/H₂O).

[0075]¹H NMR (400 MHz, CD₃OD) δ 8.08 (2H, s), 7.24 (2H, td, J=8.4 Hz,J=1.6 Hz), 7.09 (2H, dd, J=7.6 Hz, J=1.6 Hz), 7.90-7.86 (4H, m), 4.84(4H, d, J=1.6 Hz). ¹³C NMR (100 MHz, CD₃OD) δ 187.8, 158.5, 133.9,132.5, 131.9, 123.3, 120.5, 116.8, 70.0. Anal. Calcd for C₁₉H₁₆O₄: C,74.01; H, 5.23. Found: C, 73.23; H, 5.23. EIHRMS: m/z 290.0933 (M⁺,C₁₉H₁₆O₄ requires 290.0943).

[0076] 2,5-Bis(2-hydroxyphenyl)cyclopentanone (EF31): yellow solid (81%,recrystallized from acetone).

[0077]¹H NMR (400 MHz, CD₃OD) δ 8.00 (2H, s), 7.57 (2H, dd, J=8.0 Hz,J=1.2 Hz), 7.22 (2H, td, J=8.0 Hz, J=1.6 Hz), 6.88 (4H, m), 3.06 (4H,s).

[0078] 3,5-Bis(2-hydroxybenzylidene)1-methyl-4-piperidone (EF34): yellowsolid (75%, recrystallized from methanol/H₂O).

[0079]¹H NMR (400 MHz, CD₃OD) δ 8.11 (2H, s), 7.23 (4H, t, J=7.6 Hz),6.88 (4H, t, J=8.0 Hz), 3.76 (4H, d, J=1.2 Hz), 2.42 (3H, s). ¹³C NMR(100 MHz, CD₃OD) δ 188.40, 158.58, 135.09, 133.09, 132.27, 131.64,123.59, 120.38, 116.87. HREIMS: m/z 303.1259 (M⁺-H₂O, C₂₀H₁₇NO₂ requires303.1259).

Example 2 Preparation of EF8, EF9, EF10, EF23, EF29, EF30, EF33

[0080] The compounds of this series were all synthesized by thefollowing procedure: A solution of fluoro-substituted benzaldehyde (5.00mmol) in ethanol abs. (1 mL) was added at room temperature over a periodof 5 min, with stirring, to a solution of NaOH (0.75 mmol) and ketone(acetone, tetrahydro-4-H-pyran-4-one, N-methyl-piperodin-4-one) (2.50mmol) in a mixture of ethanol abs (7 mL) and H₂O dist. (7 mL). Thesolution turns yellow immediately, and usually a yellow precipitatestarts forming within 10 min (except EF8, where an oil forms). Thereaction was stirred at room temperature for 3 hrs., the yellow solidfiltered off, washed with water and hexanes and dried under vacuum. Theproduct was obtained in analytically pure form, further purification wasonly necessary where indicated. A description of each compound obtainedby the above process is given below.

[0081] 1,5-Bis(2-fluorophenyl)penta-1,4-dien-3-one (EF8): yellow solid(50%).

[0082]¹H NMR (400 MHz, CDCl_(3,)) δ 7.86 (2H, d, J=16 Hz), 7.63 (2H, td,J=7.6 Hz, J=1.6 Hz), 7.42-7.35 (2H, m), 7.18 (2H, d, J=16 Hz), 7.26-7.10(4H, m). ¹³C NMR (100 MHz, CDCl₃) δ 189.2, 161.9 (d, J=253.4 Hz), 136.3(d, J=2.3 Hz), 132.1 (d, J=9.1 Hz), 129.5 (d, J=2.2 Hz), 127.8 (d, J=6.1Hz), 124.7 (d, J=3.8 Hz), 123.1 (d, J=11.4 Hz), 116.4 (d, J=22.0 Hz).Anal. Calcd for C₁₇H₁₂F₂O: C, 75.55; H, 4.48; F, 14.06. Found: C 75.30,; H 4.55. EIHRMS: m/z 270.0865 (M⁺, C₁₇H₁₂F₂O requires 270.0856).

[0083] 1,5-Bis(2,4-difluorophenyl)penta-1,4-dien-3-one (EF9): yellowsolid (72%).

[0084]¹H NMR (300 MHz, CDCl₃) δ 7.79 (2H, d, J=16.2 Hz), 7.62 (2H, dd,J=15 Hz, J=8.4 Hz,), 7.10 (2H, d, J=16 Hz), 8.85-8.97 (4H, m). ¹C NMR(75 MHz, CDCl₃) δ 188.5, 164.8 (dd, J=169.6 Hz, J=12.2 Hz), 161.3 (dd,J=160.3 Hz, J=12.2 Hz), 135.3, 130.7 (q, J=10.0 Hz, J=4.6 Hz), 127.3,119.5, 112.3 (d, J=21.9 Hz), 104.9 (t, J=25.4 Hz). Anal. Calcd forC₁₇H₁₀F₄O: C, 66.67; H, 3.29. Found: C, 66.38; H, 3.41. EIHRMS: m/z306.0654 (M⁺, C₁₇H₁₀OF₄ requires 306.0668).

[0085] 1,5-Bis(3,4-difluorophenyl)penta-1,4-dien-3-one (EF10): yellowsolid (86%).

[0086]¹H NMR (400 MHz, CDCl₃) δ 7.64 (2H, d, J=16 Hz), 7.44 (2H, ddd,J=9.6 Hz, 7.6 Hz, J=2 Hz), 7.32-7.36 (2H, m), 7.18-7.25 (2H, m), 6.96(2H, d, J=16 Hz). ¹³C NMR (100 MHz, CDCl₃) δ 180.0, 152.7 (dd, J=105.4Hz, J=12.9 Hz), 150.2 (dd, J=101.7 Hz, J=12.9 Hz), 141.5, 132.1 (t,J=5.2 Hz), 126.2, 125.5 (q, J=6 Hz, J=3 Hz), 118.2 (d, J=17.5 Hz), 116.7(d, J=20.5 Hz). Anal. Calcd for C₁₇H₁₀F₄O: C, 66.67; H, 3.29. Found: C,66.54; H, 3.28. EIHRMS: m/z 306.0671 (M⁺, C₁₇H₁₀OF₄ requires 306.0668).

[0087] 1,5-Bis(2,6-difluorophenyl)penta-1,4-dien-3-one (EF23): yellowsolid (91%).

[0088]¹H NMR (400 MHz, CDCl₃) δ 7.82 (2H, d, J=16.4 Hz), 7.35 (2H, d,J=16 Hz), 7.33 (2H, td, J=8 Hz, J=2.4 Hz), 6.96 (4H, t, J=8.8 Hz). ¹³CNMR (100 MHz, CDCl₃) δ 189.5, 162.2 (dd, J=1017.6 Hz, J=29.2 Hz), 131.6(t, J=10.9 Hz), 131.0 (t, J=8.7 Hz), 129.9, 113.0 (t), 112.1 (d, J=25.6Hz). Anal. Calcd for C₁₇H₁₀FO₄: C, 66.67; H, 3.29. Found: C, 66.46; H,3.26, EIHRMS: m/z 306.0657 (M⁺, C₁₇H₁₀FO₄ requires 306.0668).

[0089] 3,5-Bis(2-fluorobenzylidene)tetrahydro-4-H-pyran-4-one (EF29):(84%, recrystallized from hot ethanol).

[0090]¹H NMR (400 MHz, CDDl₃) δ 7.92 (2H, d, J=1.2 Hz), 7.40-7.36 (2H,m), 7.20-7.18 (4H, m), 7.13 (2H, t, J=9.6 Hz), 4.80 (s, 4H, CH₂). ¹³CNMR (100 MHz, CDCl₃) δ 185.1, 161.0 (d, J=250.7 Hz), 134.8, 131.5 (d,J=8.7 Hz), 131.1 (d, J=2.2 Hz), 129.6 (d, J=3.7 Hz), 124.3 (d, J=3.7Hz), 122.8 (d, J=13.8 Hz), 116.2 (d, J=21.9 Hz), 68.9 (d, J=5.1 Hz).Anal. Calcd for C₁₉H₁₄F₂O₂: C, 73.07; H, 4.52. Found: C, 73.07; H, 4.47.EIHRMS; m/z 312.0950 (M⁺, C₁₉H₁₄F₂O₂ requires 312.0962).

[0091] 3,5-Bis(2,4-difluorobenzylidene)tetrahydro-4-H-pyran-4-one(EF30): (82%, recryst. from ethanol/H₂O).

[0092]¹H NMR (400 MHz, CDCl₃) δ 7.85 (2H, s), 7.21-7.14 (1H, m),6.98-6.87 (2H, m), 4.77 (4H, s). 13C NMR (100 MHz, CDCl₃) δ 184.81,163.95 (dd, J=240 Hz, J=12 Hz), 161.43 (dd, J=265 Hz, J=12 Hz), 134.48,132.02 (dd, J=9.5 Hz, J=4.3 Hz), 128.62, 119.19, 112.00 (dd, J=3.7 Hz,J=21.9 Hz), 104.85 (t, J=25.5 Hz), 68.81 (d, J=4.4 Hz). Anal. Calcd forC₁₉H₁₂F₄O₂; C, 65.52; H, 3.47. Found: C, 65.67; H, 3.43. EIHRMS: m/z348.0761 (M⁺, C₁₉H₁₄F₂O₂ requires 348.0773).

[0093] 3,5-Bis(2-fluorobenzylidene)1-methyl-4-piperidone (EF33): (82 %,yellow solid).

[0094]¹H NMR (400 MHz, CDCl₃) δ 7.90 (2H, s), 7.40-7.33 (2H, m), 7.28(2H, td, J=7.6 Hz, J=1.2 Hz), 7.18 (2H, td, J=7.6 Hz, J=0.8 Hz), 7.12(2H, td, J=10.0 Hz, J=0.8 Hz), 3.64 (4H, s), 2.40 (3H, s). ¹³C NMR (100MHz, CDCl₃) δ 186.3, 161.1 (d, J=251 Hz), 134.9, 131.1 (d, J=8.1 Hz),131.0 (d, J=3.0 Hz), 129.7 (d, J=3.6 Hz), 124.1 (d, J=2.9 Hz), 123.4 (d,J=13.9 Hz), 116.1 (d, J=21.8 Hz), 57.2, 45.8. Anal. Calcd forC₂₀H₁₇F₂ON: C, 73.83; H, 5.27; N, 4.30. Found: C, 73.59; H, 5.32; N,4.39. EIHRMS: m/z 325.1278 (M⁺, C₂₀H₁₇F₂ON requires 325.1278).

Example 3 Preparation of EF11, EF12, EF13, EF14, EF15

[0095] The compounds of this series were all synthesized by thefollowing procedure: Bis-diethylphosphorylmethylsulfide, -sulfoxide and-sulfone were obtained according to literature procedures (Tetrahedron,1992, 48, 8065-8072; Phosphorus Sulfur 1981, 10, 369-374). A solution ofphosphonate (0.60 mmol) and aldehyde (1.25 mmol) in CH₂Cl₂ (3 mL) wasadded to the heterogeneous mixture of 50 % aqueous NaOH (2 mL) andCH₂Cl₂ (2 mL), containing triethylbenzylammonium chloride (TEBA, 0.06mmol). The reaction was stirred at room temperature over night, theproduct was extracted from the reaction mixture with CH₂Cl₂ and purifiedby column chromatography. A description of each compound obtained by theabove process is given below.

[0096] 3,5-Bis(2-hydroxybenzylidene)-sulfone (EF11): yellow solid (45%,30% EtOAc/hexanes).

[0097]¹H NMR (400 MHz, CD₃OD) δ 7.77 (2H, d, J=15.6 Hz), 7.48 (2H, dd,J=8.4 Hz, J=2.0 Hz), 7.25 (2H, td, J=8.0 Hz, J=2.5 Hz), 7.22 (2H, d,J=15.6 Hz), 6.88-6.84 (4H, m), 4.91 (2H, s-br). ¹³C NMR (100 MHz, CD₃OD)δ 158.94, 140.21, 133.55, 131.63, 127.84, 121.15, 121.03, 117.23.

[0098]3,5-Bisbenzylidenesulfone (EF12): white solid (78%, 20% EtOAc/hexanes).

[0099]¹H NMR (400 MHz, CDCl₃) δ 7.65 (2H, d, J=15.2 Hz, H3), 7.53-7.50(4H, m), 7.44-7.40 (6H m), 6.86 (2H, d, J=15.6 Hz, H2). ¹³C NMR (100MHz, CDCl₃) δ 143.7, 132.7, 131.5, 129.3, 128.8, 126.5. Anal. Calcd forC₁₆H₁₄SO₂: C, 71.09; H, 5.22, S, 11.86. Found: C, 70.88; H, 5.21, S,12.01. EIHRMS: m/z 270.0715 (M⁺, C₁₆H₁₄SO2 requires 270.0731).

[0100] E,E-3,5-Bisbenzylidenesulfoxide (EF13): white solid (33%, 20%EtOAc, hexanes).

[0101]¹H NMR (300 MHz, CDCl₃) δ 7.52-7.46 (4H, m), 7.42-7.35 (6H, m),7.31 (2H, d, J=15.3 Hz), 6.87 (2H, d, J=15.3 Hz). ¹³NMR (75 MHz, CDCl₃)δ 137.0, 133.9, 131.1, 130.0, 129.1, 127.9. Anal. Calcd for C₁₆H₁₄SO: C,75.56; H, 5.55, S, 12.60. Found: C, 75.33; H, 5.60, S, 12.60.

[0102] E,Z-3,5-Bisbenzylidenesulfoxide (EF14): white solid (15%, 20%EtOAc, hexanes).

[0103]¹H NMR (300 MHz, CDCl₃) δ 7.52-7.40 (10H, m), 7.33 (1H, d, J=15.3Hz), 7.12 (1H, d, J=10.5 Hz), 6.93 (1H, d, J=15.6 Hz), 6.39 (1H, d,J=10.5 Hz). ¹³C NMR (75 MHz, CDCl₃) δ 138.5, 136.3, 135.5, 134.1, 130.1,129.8, 129.6, 129.0, 128.8, 127.8. Anal. Calcd for C16H₁₄SO: C, 75.56;H, 5.55, S, 12.60. Found: C, 75.34; H, 5.54, S, 12.57.

[0104]3,5-Bisbenzylidenesulfide (EF15): white solid (20%, 5%EtOAc/hexanes) mixture of E,E and E,Z (ca. 2.5:1).

[0105]¹H NMR (400 MHz, CDCl₃) δ 7.40-7.10 (20H, m), 6.79 (2H, d, J=15.6Hz), 6.76 (1H, d, J=15.6 Hz), 6.62 (2H, d, J=15.6 Hz), 6.61(1H, d,J=15.6 Hz), 6.53 (1H, J=10.8 Hz), 6.41 (1H, d, J=10.8 Hz). Anal. Calcdfor C₁₆H₁₄S: C, 80.63; H, 5.92; S, 13.45. Found: C, 80.40; H, 5.93; S,13.39.

Example 4 Preparation of EF16, EF17, EF18, EF27, EF28

[0106] The compounds of this series were all synthesized by thefollowing procedure: NaOH (0.10 mmol) was added as a solid to a stirredsolution of methoxy-substituted benzaldehyde/anisaldehyde (2.50 mmol)and ketone (acetone, tetrahydro-4-H-pyranone) in EtOH abs (5 mL). Ayellow solid started forming within 1 hr. The reaction was stirred atroom temperature for 20 hrs, the product filtered off, washed with coldEtOH abs and H₂O dist and dried under vacuum. A description of eachcompound obtained by the above process is given below.

[0107] 1,5-Bis(2-methoxphenyl)penta-1,4-dien-3-one (EF16): yellow solid(60%). Mp 123-124 ° C. (EtOH).

[0108]¹H NMR (400 MHz, CDCl₃) δ 8.07 (2H, d, J=16 Hz,), 7.63 (2H, dd,J=7.6 Hz, J=1.6 Hz), 7.37 (2H, ddd, J=7.2 Hz, J=1.6 Hz, J=1.2 Hz), 7.18(2H, d, J=16 Hz), 6.99 (2H, t, J=7.6 Hz), 6.93 (2H, d, J=8.4 Hz). ¹³CNMR (100 MHz, CDCl₃) δ 190.2, 158.7, 138.4, 131.8, 128.9, 126.4, 124.1,120.9, 111.4, 55.7. Anal. Calcd for C₁₉H₁₈O₃: C, 77.53; H, 6.16. Found:C, 77.26; H, 6.17. EIHRMS: m/z 294.1256 (M⁺, C₁₉H₁₈O₃ requires294.1256).

[0109] 1,5-Bis(3-methoxyphenyl)penta-1,4-dien-3-one (EF17): yellow solid(40%, chromatography using 20 % EtOAc/hexanes).

[0110]¹H NMR (300 MHz, CDCl₃) δ 7.70 (2H, d, J=15.9 Hz), 7.33 (2H, t,J=7.6 Hz), 7.21 (2H, d, J=7.8 Hz), 7.13 (2H, t, J=2.4 Hz), 7.06 (2H, d,J=15.9 Hz), 6.96 (2H, ddd, J=8.1 Hz, J=2.4 Hz, J=0.9 Hz). ¹³C NMR (75MHz, CDCl₃) δ 188.83, 159.96, 143.31, 136.21, 130.03, 125.72, 121.20,116.46, 113.36, 55.55.

[0111]1,5-Bis(4-methoxyphenyl)penta-1,4-dien-3-one (EF18): yellow solid(93%). Mp 129-130 ° C. (EtOH).

[0112]¹H NMR (300 MHz, CDCl₃) δ 7.69 (2H, d, J=15.9 Hz, 3), 7.56 (4H, d,J=9 Hz), 6.95 (2H, d, J=15.6 Hz), 6.92 (4H, d, J=8.7 Hz). ¹³C NMR (75MHz, CDCl₃) δ 188.8, 161.5, 142.7, 130.2, 127.7, 126.6, 114.5, 55.6Anal. Calcd for C₁₉H₁₈O₃: C, 77.53; H, 6.16. Found: C, 77.31; H, 6.217.EIHRMS: m/z 294.1268 (M⁺, C,₁₉H₁₈O₃ requires 294.1256).

[0113] 3,5-Bis(4-methoxybenzylidene)tetrahydro-4-H-pyran-4-one (EF27):yellow solid (40%).

[0114]¹H NMR (400 MHz, CDCl₃) δ 7.79 (2H, s), 7.29 (4H, d, J=8.8 Hz,),6.95 (4H, d, J=9.2 Hz), 4.93 (4H, d, J=1.6 Hz), 3.85 (6H, s). ¹³C NMR(100 MHz, CDCl₃) δ 185.6, 160.8, 136.2, 132.7, 131.4, 127.7, 114.4,68.9, 55.6. Anal. Calcd for C₂₁H₂₀O₄: C, 74.98; H, 5.99. Found: C,74.81; H, 6.01. EIHRMS: m/z 336.1362 (M⁺, C₂₁H₂₀O₄ requires 336.1361).

[0115] 3,5-Bis(2-methoxybenzylidene)tetrahydro-4-H-pyran-4-one (EF28):yellow solid (67%).

[0116]¹H NMR (400 MHz, CDCl₃) δ 8.09 (2H, s), 7.36 (2H, td, J=8.8 Hz,J=1.6 Hz), 7.07 (2H, dd, J=7.6 Hz, J=1.6 Hz), 6.97 (2H, t, J=7.6 Hz),6.93 (2H, d, J=8 Hz), 4.81 (4H, d, J=1.6 Hz), 3.87 (6H, s). ¹³C NMR (100MHz, CDCl₃) δ 186.0, 158.6, 133.3, 132.5, 131.1, 130.7, 124.1, 120.3,111.0, 69.1, 55.7. Anal. Calcd for C₁₂H₂₀O₄: C, 74.98; H, 5.99. Found:C, 74.84; H, 5.92. EIHRMS: m/z 336.1370) (M⁺, C₂₁H₂₀O₄ requires336.1361).

Example 5 Preparation of EF19, EF20, EF32

[0117] The compounds of this series were all synthesized by thefollowing procedure: A solution of substituted dienone (0.69 mmol) inEtOH abs (29 mL) was subject to hydrogenation at 33 psi using RaneyNickel as the catalyst for 4 hrs. Filtration through CELITE andconcentration under vacuum yielded the crude product, which was purifiedby chromatography on silica gel using 25% EtOAc/hexanes. A descriptionof each compound obtained by the above process is given below.

[0118] 1,5-Bis(2,4-difluorophenyl)-pentan-3-ol (EF19): white solid(92%).

[0119]¹H NMR (300 MHz, CDCl₃) δ 7.17-7.09 (2H, m), 6.82-6.72 (4H, m),3.60 (1H, m), 2.84-2.60 (4H, m), 1.80-1.75 (4H, m), 1.59 (1H, s-br). 13CNMR (75 MHz, CDCl₃) δ 162.9 (dd, J=34.4 Hz, J=12 Hz), 159.7 (dd, J=35Hz, J=12 Hz), 131.1 (t, J=9 Hz), 124.6 (dd, J=19.7 Hz, J=3.8 Hz), 111.2(dd, J=20.5 Hz, J=3.4 Hz), 103.8 (t, J=25.7 Hz), 70.6, 38.0, 25.1. Anal.Calcd for C₁₇H₁₆FO₄: C, 65.38; H, 5.16. Found: C, 65.64; H, 5.24.EIHRMS: m/z 312.1137 (M⁺, C₁₇H₁₆FO₄ requires 312.1137).

[0120] 1,5-Bis(3,4-difluorophenyl)-pentan-3-ol (EF20): white solid(81%).

[0121]¹H NMR (300 MHz, CDCl₃) δ 7.07-6.91 (4H, m), 6.87-6.80 (2H, m),3.57 (1H, m), 2.77-2.54 (4H, m), 1.73 (4H, t), 1.41 (1H, s-br). ¹³C NMR(75 MHz, CDCl₃) δ 151.2 (dd, J=107.6 Hz, J=12.5 Hz), 148.0 (dd, J=105.8Hz, J=12.5 Hz), 138.9 (t), 124.3 (d, J=5.8 Hz), 117.3, 117.1, 70.4,39.3, 31.5. EIHRMS: m/z 312.1138(M⁺, Cl₇H₁₆FO₄ requires312.1137).

[0122] 1,5-Bis(2-hydroxyphenyl)-pentan-3-ol (EF32): white solid (45%).

[0123]¹H NMR (300 MHz, CDCl₃) δ 7.56 (2H, s-br), 7.09-7.05 (4H, m),6.87-6.80 (4H, m), 3.54 (1H, m), 2.92-2.84 (2H, m), 2.69-2.62 (2H, m),1.80-1.71 (4H, m). ¹³C NMR (75 MHz, CDCl₃) δ 154.11, 130.84, 127.91,127.77, 121.14, 116.05, 69.28, 37.89, 25.87.

Example 6 Preparation of EF21, EF22

[0124] The compounds of this series were all synthesized by thefollowing procedure: PCC (107 mg, 0.50 mmol) was added in one portion atroom temperature to a stirred solution of alcohol (103 mg. 0.33 mmol) inCH₂Cl₂ (5mL). The reaction was stirred at room temperature for 14 hrs(TLC-analysis, no sm left), filtered over CELITE and concentrated. Thecrude product was purified by chromatography on silica gel using 20%EtOAc/hexanes. A description of each compound obtained by the aboveprocess is given below.

[0125] 1,5-Bis(2,4-difluorophenyl)-pentan-3-on (EF21): white solid(92%).

[0126]¹H NMR (400 MHz, CDCl₃) δ 7.16-7.09 (2H, m), 6.80-6.72 (4H, m),2.87 (4H, t, J=7.6 Hz), 2.69 (4H, t, J=7.6 Hz). ¹³C NMR (100 MHz, CDCl₃)δ 208.08, 163.00 (dd, J=48 Hz, J=11.7 Hz), 159.73 (dd, J=49 Hz, J=12Hz), 131.39, 123.54 (d, J=15.9), 111.17 (dd, J=20.8 Hz, J=3.8 Hz),103.84 (t, J=25.5 Hz), 42.93, 23.08.

[0127]1,5-Bis(3,4-difluorophenyl)-pentan-3-on (EF22): white solid (86%).

[0128]¹H NMR (400 MHz, CDCl₃) δ 7.07-7.00 (2H, ddd), 6.98-6.92 (2H, m),6.87-6.80 (2H, m), 2.84 (4H, t, J=7.2 Hz), 2.68 (4H, t, J=7.2 Hz).

Example 7 Preparation of EF7

[0129] EF7 was obtained in a three-step synthesis. To a solution of2,5-bis(2-hydroxybenzylidene)acetone (800 mg. 3.00 mmol) in DMF (10 mL)was added imidazole (545 mg, 7.56 mmol) and DMAP (10 mg). The brightyellow solution was cooled to 0° C. and tbutyldiphenylchlorosilane (1.75mL, 6.73 mmol) was added dropwise. After stirring for 30 minutes thecooling bath was removed, and the reaction proceeded at room temperatureuntil no more starting material or monoprotected alcohol were detectableby TLC. (hexanes/EtOAc=2/1, Rf (starting material)=0.17, Rf (mono)=0.44,Rf (di)=0.78). The orange solution was poured into ice water (50 mL) andextracted with ether (3×). The combined organic layers were washed withbrine (3×), dried over MgSO₄ and concentrated. The crude product waspurified by plug chromatography on silica gel (15% EtOAc/hexanes). Theproduct (108 %) was obtained as a yellow foam and contains sometbutyldiphenylsilylalcohol. It was used without further purification.

[0130]¹H NMR (400 MHz, CDCl₃) δ 8.48 (2H, d, J=16.0 Hz), 7.75-7.71 (8H,m), 7.66 (2H, dd, J=8.0 Hz, J=2.0 Hz), 7.46-7.35 (12H, m), 7.18 (2H, d,J=16.4 Hz), 6.96 (2H, td), 6.89 (2H, td), 6.50 (2H, dd, J=8.0 Hz, J=1.2Hz), 1.08 (18H, s). ¹³C NMR (100 MHz, CDCl₃) δ 189.93, 154.83, 138.24,135.63, 135.38, 132.35, 130.30, 128.14, 127.92, 127.26, 125.43, 121.54,120.08, 26.81, 19.88.

[0131] Disilylprotected ketone (1.74 g, 2.34 mmol) was dissolved in THFand cooled to −78° C., where CH₃Li (1.90 mL, 2.66 mmol, 1.4 M/ether) wasadded dropwise. After stirring for 10 min the originally bright yellowsolution completely cleared, it was quenched with saturated NH₄Cl, thelayers separated and the aqueous phase extracted with ether. Thecombined organic layers were washed with brine, dried over MgSO₄ andconcentrated. The crude product was purified by chromatography on silicagel (10% EtOAc/hexanes). 60%, white foam.

[0132]¹H NMR (400 MHz, CDCl₃) δ 7.72 (8H, dd, J=6.4 Hz, J=1.6 Hz), 7.53(2H, dd, J=7.2 Hz, J=2.4 Hz), 7.41 (4H, m), 7.36-7.32 (10H, m), 6.82(4H, m), 6.45 (2H, d, J=16.4 Hz), 6.43 (2H, dd, J=7.2 Hz, J=2.8 Hz),1.67 (3H, s), 1.09 (18H, s). ¹³C NMR (100 MHz, CDCl₃) δ 152.96, 135.66,135.33, 132.81, 130.12, 128.33, 128.01, 127.72, 126.42, 123.37, 121.37,119.62, 74.02, 28.39, 26.73, 19.79.

[0133] EF7: Deprotection of the alcohol was carried out withtetrabutylammonium fluoride (2.2 equiv) in THF. The product was obtainedafter column chromatography on silica gel (30% EtOAc/hexanes) as a whitesolid (48 %).

[0134]¹H NMR (400 MHz, CDCl₃) δ 7.32 (1H, dd, J=7.6 Hz, J=1.6 Hz),7.14-7.07 (2H, m), 7.00 (1H, dd, J=8.0 Hz, J=1.6 Hz), 6.89-6.82 (3H, m),6.75 (1H, dd, J=8 Hz, J=1.2 Hz), 6.47 (1H, d, J=9.6 Hz), 6.31 (1H, d,J=16.4 Hz), 5.69 (1H, d, J=10 Hz), 5.00 (1H, s-br), 1.66 (3H, s). ¹³CNMR (100 MHz, CDCl₃) δ 153.1, 153.0, 133.9, 129.5, 129.0, 128.1, 127.8,126.7, 123.9, 123.7, 121.4, 121.2, 121.1, 116.5, 116.0, 78.0, 27.6.Anal. Calcd for C₁₈H₁₆O₂ C, 81.79; H, 6.10. Found: C, 81.61; H, 6.16.EIHRMS: m/z 264.1150 (M⁺, C₁₈H₁₆O₂ requires 264.1150).

Example 8 Preparation of EF5

[0135]1,5-Bis(2-hydroxyphenyl)penta-1-en-3-ol (EF5): To a solution of1,5-bis(2-hydroxyphenyl)penta-1,4-dien-3-one (EF1) (109 mg, 0.41 mmol)in THF/methanol (10/1) (2.5 mL) was added NaBH₄ (40mg, 1.30 mmol) in oneportion at 0C. After stirring for 30 min at this temperature thereaction was quenched with H₂O dist and cold brine, diluted with Et₂O(10 mL) and neutralized by bubbling CO₂ through the dark-red solution(color change to pale yellow). The aqueous phase was extracted withether, the combined ether layers washed with brine, dried over MgSO₄ andconcentrated. The crude product was purified by chromatography on silicagel using 30% EtOAc/hexanes. The product was obtained as a white solid(66%).

[0136]¹H NMR (300 MHz, CDCl₃) δ 7.27 (2H, dd, J=8.0 Hz, J=2.4 Hz), 7.21(1H, s-br), 7.14-7.06 (3H, m), 6.90-6.83 (3H, m), 6.80-6.74 (2H, m),6.18 (1H, dd, J=20.8 Hz, J=8.8 Hz), 5.99 (1H, s-br), 4.24 (1H,t-deformed), 3.23 (1H, s-br), 2.86 (1H, m), 2.71 (1H, m), 1.91 (2H, m).¹³C NMR (75 MHz, CDCl₃) δ 154.25, 152.96, 132.84, 130.75, 129.03,127.79, 127.62, 127.46, 125.71, 123.97, 121.12, 121.07, 116.42, 116.22,72.09, 37.57, 25.76.

Example 9 Preparation of EF6

[0137] N-(Methoxy)-1,5-bis(2-hydroxyphenyl)penta-1,4-dien-3-imine (EF6):Methoxyl-amine hydrochloride (30-35 wt % in H₂O, 0.30 mL, 1.12 mmol) wasadded in one portion to a solution of1,5-bis(2-hydroxyphenyl)penta-1,4-dien-3-one (EF1) in methanol/CHCl₃(2/3) (5 mL). The reaction proceeded at room temperature for 24 hrs,then additional methoxylamine hydrochloride (0.15 mL, 0.56 mmol) wasadded and the reaction stirred for an additional 24 hrs. Aftercompletion (TLC) the solvent was evaporated, the residue dissolved inmethanol, stirred with silica gel and purified after concentration bychromatography on silica gel using 30% EtOAc/hexanes. The product wasobtained as a pale yellow foam (86%).

[0138]¹H NMR (400 MHz, CDCl₃) δ 7.56 (1H, dd, J=8.0 Hz, J=1.6 Hz), 7.48(1H, dd, J=8.0 Hz, J=1.6 Hz), 7.41 (1H, d, J=16.0 Hz), 7.39 (1H, d,J=16.8 Hz), 7.28 (1H, d, J=16.8 Hz), 7.20-7.13 (2H, m), 6.97-6.90 (2H,m), 6.94 (1H, d, J=16.0 Hz), 6.80 (1H, dd, J=8.0 Hz, J=1.6 Hz), 5.53(1H, s-br), 5.36 (1H, s-br), 4.02 (3H, s). ¹³C NMR (100 MHz, CDCl₃) δ155.43, 153.86, 153.70, 132.28, 130.25, 130.10, 129.71, 128.00, 127.91,124.16, 123.88, 123.57, 121.40, 121.30, 118.45, 116.37, 116.33, 62.41.

Example 10 Preparation of MD279U and MD279L

[0139] A solution of 1,5-bis(3,4-dimethoxyphenyl)penta-1,3-dien-3-one ina mixture of EtOH abs and THF (5/1) was subject to hydrogenation at 50psi using Raney Nickel as the catalyst for 8 hrs. Filtration throughCELITE and concentration under vacuum yielded the crude product, whichwas purified by chromatography on silica gel using 25% EtOAc/hexanes.

[0140] 1,5-Bis(3,4-dimethoxyphenyl)penta-3-one (MD279U): 37%, whitesolid.

[0141]¹H NMR (400 MHz, CDCl₃) δ 6.77 (2H, d, J=8.8 Hz), 6.69 (4H, m),3.86 (6H, s), 3.85 (6H, s), 2.84 (4H, t, J=7.6 Hz), 2.70 (4H, t, J=7.6Hz). ¹³C NMR (100 MHz, CDCl₃) δ 209.61, 149.00, 147.50, 133.77, 120.22,111.82, 111.38, 56.07, 55.98, 45.01, 29.54. R_(f)=0.28(EtOAc/hexanes=½).

[0142] 1,5-Bis(3,4-dimethoxyphenyl)penta-3-ol (MD279L): 51%, whitesolid.

[0143]¹H NMR (400 MHz, CDCl₃) δ 6.79 (2H, d, J=8.8 Hz), 6.73 (4H, m),3.87 (6H, s), 3.86 (6H, s), 3.68 (1H, m), 2.78-2.71 (2H, m), 2.63 (2H,ddd, J=14.0 Hz, J=9.2 Hz, J=6.8 Hz), 1.79 (4H, m), 1.58 (1H, s-br). ¹³CNMR (100 MHz, CDCl₃) δ 149.02, 147.34, 134.80, 120.29, 111.84, 111.39,71.04.

Example 11 3,5-Bis-(α,α,α-trifluoro-2-toluylbenzylidene)-piperidin-4-oneacetate

[0144]

[0145] A suspension of 1.76 g (11.49 mmol) of 4-piperidone hydrate, HClsalt, in 60.0 mL of glacial acetic acid was saturated with dry HCl gasand to the resultant solution was added 5.0 g (28.72 mmol) ofα,α,α-trifluoro-2-tolualdehyde. The mixture was allowed to stir at roomtemperature for 72 hrs and then diluted with 50.0 mL of toluene andevaporated under vacuum. The residue was diluted twice more with 50.0 mLportions of toluene and evaporated under vacuum. The gummy residue wassuspended in 50 mL of toluene containing 5.0 mL of ethyl acetate, heatedbriefly to reflux and allowed to cool to room temperature. The solidsformed were collected by suction filtration and dried under high vacuumto afford 3.63 g (67%)of a bright yellow solid.

Example 12 3-5-Bis-(pyridinylidene)-piperidin-4-one

[0146]

[0147] To a solution of 1.00 g (6.52 mmol) of 4-piperidone hydrate, HClsalt, and 1.40 g (13.05 mmol) of 2-pyridine carboxaldehyde in 91 mL of a0.25M solution of aqueous NaOH (22.82 mmol) was added 0.86 mL of a 25%w/w solution (0.65 mmol) of cetyltrimethylammonium chloride. The mixturewas allowed to stir vigorously at room temperature for three hours,diluted with 100 ml of brine and extracted with three 50 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO4 andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto afford 1.25g (69%) of a red-yellow solid.

Example 133,5-Bis-(2-fluoro-3-α,α,α-trifluoromethylbenzylidene)-piperidin-4-one

[0148]

[0149] To a solution of 400 mg (2.61 mmol) of 4-piperidone hydrate, HClsalt, and 1.00 g (5.21 mmol) of2-fluoro-3-α,α,α-trifluoromethylbenzaldehyde in 36 mL of a 0.25Msolution of aqueous NaOH (9.11 mmol) was added 0.35 mL of a 25% w/wsolution (0.26 mmol) of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 48 hours, dilutedwith 100 ml of brine and extracted with two 50 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo to afford 1.02 g (86%) of a yellow foam.

Example 14

[0150]

[0151] To a solution of 1.0 g (6.52 mmol) of 4-piperidone hydrate, HClsalt, and 1.88 g (13.37 mmol) of 2-chloro benzaldehyde in 95 mL of a0.25M solution of aqueous NaOH (22.82 mmol) was added 0.90 mL of a 25%w/w aqueous solution (0.65 mmol) of cetyltrimethylammonium chloride. Themixture was allowed to stir vigorously at room temperature for 48 hours,diluted with 100 ml of brine and extracted with two 35 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto give 0.92 g (41%) of a pale yellow powder.

Example 15

[0152]

[0153] To a solution of 1.0 g (8.84 mmol) of 1-methylpiperidine-4-oneand 3.08 g (13.37 mmol) of α,α,α-trifluoro-2-tolualdehyde in 88 mL of a0.25M solution of aqueous NaOH (22.09 mmol) was added 1.16 mL (0.88mmol) of a 25% w/w aqueous solution of cetyltrimethylammonium chloride.The mixture was allowed to stir vigorously at room temperature for 3hours, diluted with 100 ml of brine and extracted with three 50 mLportions of methylene chloride. The organic phase was dried overanhydrous MgSO₄ and concentrated in vacuo to give 3.56 g (95%) of a paleyellow powder.

Example 16 3,5-Bis-(2-pyrilidinyldene)-1-methylpiperidin-4-one

[0154]

[0155] To a solution of 1.0 g (8.84 mmol) of 1-methylpiperidine-4-oneand 1.89 g (16.70 mmol) of 2-pyridine carboxaldehyde in 88 mL of a 0.25Msolution of aqueous NaOH (22.09 mmol) was added 1.16 mL (0.88 mmol) of a25% w/w aqueous solution of cetyltrimethylammonium chloride. The mixturewas allowed to stir vigorously at room temperature for 3 hours, dilutedwith 100 ml of brine and extracted with three 50 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo to give 2.50 g (97%) of a pale yellow powder.

Example 17 3,5-Bis-(4-pyridinylidene)-1-methylpiperidin-4-one

[0156]

[0157] To a solution of 1.0 g (8.84 mmol) of 1-methylpiperidine-4-oneand 1.89 g (16.70 mmol) of 4-pyridine carboxaldehyde in 88 mL of a 0.25Msolution of aqueous NaOH (22.09 mmol) was added 1.16 mL (0.88 mmol) of a25% w/w aqueous solution of cetyltrimethylammonium chloride. The mixturewas allowed to stir vigorously at room temperature for 3 hours, dilutedwith 100 ml of brine and extracted with two 60 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo to give 2.15 g (84%) of a yellow-orange powder.

Example 18 3,5-Bis-(2,6-difluorobenzylidene)-1-methylpiperidin-4-one

[0158]

[0159] To a solution of 1.0 g (8.84 mmol) of 1-methylpiperidine-4-oneand 2.57 g (18.09 mmol) of 2,6-difluoro benzaldehyde in 90 mL of a 0.25Msolution of aqueous NaOH (22.59 mmol) was added 1.16 mL (0.88 mmol) of a25% w/w aqueous solution of cetyltrimethylammonium chloride. The mixturewas allowed to stir vigorously at room temperature for 12 hours, dilutedwith 100 ml of brine and extracted with three 40 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was slurried in 100 ml of boilingethyl acetate and the insolubles were removed by rapid suctionfiltration. The filtrate was concentrated in vacuo and recrystallizedfrom ethyl acetate to provide 3.01 g (94%) of a bright yellow solid.

Example 19 3,5-Bis-(2,6-difluorobenzylidene)-tropin-4-one

[0160]

[0161] To a solution of 0.50 g (3.59 mmol) of tropinone and 1.05 g (7.39mmol) of 2,6-difluoro benzaldehyde in 36 mL of a 0.25M solution ofaqueous NaOH (9.04 mmol) was added 0.71 mL (0.54 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 4 hours, diluted with100 ml of brine and extracted with two 25 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 840 mg (60%) of a bright yellow solid.

Example 20 3,5-Bis-(2-fluorobenzylidene)-tropin-4-one

[0162]

[0163] To a solution of 0.50 g (3.59 mmol) of tropinone and 0.914 g(7.36 mmol) of 2-fluoro benzaldehyde in 36 mL of a 0.25M solution ofaqueous NaOH (9.04 mmol) was added 0.47 mL (0.36 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 12 hours, dilutedwith 100 ml of brine and extracted with two 50 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide

Example 21 3,5-Bis-(2-pyridinylidene)-tropin-4-one

[0164]

[0165] To a solution of 0.75 g (5.39 mmol) of tropinone and 1. 18 g(7.39 mmol) of 2-pyridine carboxaldehyde in 54 mL of a 0.25M solution ofaqueous NaOH (13.47 mmol) was added 0.71 mL (0.54 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 48 hours, dilutedwith 150 ml of brine and extracted with three 50 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 180 mg (11%) of a tan powder.

Example 22 1,5-Bis-(2,3-dimethyoxyphenyl)-penta-1,4-dien-3-one

[0166]

[0167] To a solution of 0.5 g (8.61 mmol) of acetone and 2.86 g (17.22mmol) of 2,3-dimethoxy benzaldehyde in 86 mL of a 0.25M solution ofaqueous NaOH (21.59 mmol) was added 2.83 mL (2.15 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 72 hours, dilutedwith 100 ml of brine and extracted with three 50 mL portions ofmethylene chloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 1.99 g (65%) of a bright yellow solid.

Example 23 1,5-Bis-(2,3-methylenedioxyphenyl)-penta-1,4-dien-3-one

[0168]

[0169] To a solution of 0.19 g (3.27 mmol) of acetone and 1.00 g (6.66mmol) of 2,3-methylenedioxy benzaldehyde in 35 mL of a 0.25M solution ofaqueous NaOH (8.79 mmol) was added 0.65 mL (0.49 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 2 hours, at whichpoint it was diluted with 15 mL of 95% ethanol, and stirring wascontinued for an additional 2 hours. The solution was saturated withsodium chloride and extracted with two 35 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 1.03 g (98%) of a yellow solid.

Example 24 1,5-Bis-(4-dimethylaminophenyl)-penta-1,4-dien-3-one

[0170]

[0171] To a solution of 1.0 g (17.22 mmol) of acetone and 5.26 g (35.30mmol) of 4-dimethylamino benzaldehyde in 172 mL of a 0.25M solution ofaqueous NaOH (43.05 mmol) was added 2.26 mL (1.72 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 72 hours, dilutedwith 100 ml of brine and extracted with two 75 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 1.21 g (22%) of a dark red powder.

Example 25 1,5-Bis-(2,6-dimethoxyphenyl)-penta-1,4-dien-3-one

[0172]

[0173] To a solution of 0.25 g (4.31 mmol) of acetone and 1.47 g (8.85mmol) of 2,6-dimethoxy benzaldehyde in 43 mL of a 0.25M solution ofaqueous NaOH (10.80 mmol) was added 0.57 mL (0.43 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 72 hours, dilutedwith 100 ml of brine and extracted with two 75 mL portions of methylenechloride. The organic phase was dried over anhydrous MgSO₄ andconcentrated in vacuo. The residue was recrystallized from ethyl acetateto provide 1.10 g (72%) of a yellow powder.

Example 26 1,5-Bis-(2,3-difluorophenyl)-penta-1,4-dien-3-one

[0174]

[0175] To a solution of 0.5 g (8.61 mmol) of acetone and 2.51 g (17.65mmol) of 2,3-difluoro benzaldehyde in 86 mL of a 0.25M solution ofaqueous NaOH (21.52 mmol) was added 1.13 mL (0.86 mmol) of a 25% w/waqueous solution of cetyltrimethylammonium chloride. The mixture wasallowed to stir vigorously at room temperature for 48 hours, at whichpoint it was diluted with 100 ml of brine and extracted with two 75 mLportions of methylene chloride. The organic phase was dried overanhydrous MgSO₄ and concentrated in vacuo. The residue wasrecrystallized from ethyl acetate to provide 110 mg (4%) of a yellowpowder.

Example 27 (E)-3-(2-fluorobenzylidenyl)indolin-2-one

[0176]

[0177] To a solution containing 2.0 g (15.02 mmol) of 2-oxindole and2.05 g (16.52 mmol) of 2-fluoro benzaldehyde in 30 mL of abs. ethanolwas added 190 mg (2.25 mmol) piperidine and the mixture was refluxed for12 hrs. The mixture was allowed to cool to room temperature and thesolids formed were collected by suction filtration and washed with two25 mL portions of cold abs. ethanol. The recovered material was driedunder high vacuum for 12 hrs. to afford 3.22 g (90%) of a bright yellowpowder.

Example 28 (E)-3-(2-pyridinylidenyl)indolin-2-one

[0178]

[0179] To a solution containing 2.0 g (15.02 mmol) of 2-oxindole and1.77 g (16.52 mmol) of 2-pyridine carboxaldehyde in 30 mL of abs.Ethanol was added 192 mg (2.25 mmol) piperidine and the mixture wasrefluxed for 12 hrs. The mixture was allowed to cool to room temperatureand the solids formed were collected by suction filtration and washedwith two 25 mL portions of cold abs. ethanol. The recovered material wasdried under high vacuum for 12 hrs to afford 2.82 g (84%) of a pale redpowder.

Example 29 (E)-3-(2,3-difluorobenzylidenyl)indolin-2-one

[0180]

[0181] To a solution containing 2.0 g (15.02 mmol) of 2-oxindole and2.20 g (16.52 mmol) of 2,3-difluoro benzaldehyde in 30 mL of abs.ethanol was added 192 mg (2.25 mmol) piperidine and the mixture wasrefluxed for 12 hrs. The mixture was allowed to cool to room temperatureand the solids formed were collected by suction filtration and washedwith two 25 mL portions of cold abs. ethanol. The recovered material wasdried under high vacuum for 12 hrs.

Example 30 1,3-Bis-(2-fluorobenzylidene)indan-2-one

[0182]

[0183] A solution of 1.88 g (15.13 mmol) of 2-fluorobenzaldehyde in 3.0mL of abs. ethanol was added over a period of 5 min to a solutioncontaining 1.00 g (7.57 mmol) of 2-indanone and 90 mg (2.27 mmol) ofNaOH in 40 mL of a 1:1 mixture of abs. ethanol and water at roomtemperature. The mixture was allowed to stir for 12 hrs and solidsformed were collected by suction filtration and washed with cold ethanoland dried under high vacuum.

Example 31 3,5-Bis-(2-fluorobenzylidene)-piperidin-4-one-acetate-EF24

[0184]4-Piperidone hydrochloride monohydrate (307 mg, 2.00 mmol) wassuspended in glacial acetic acid (8 mL) and saturated with HCl gas atroom temperature. To the resulting clear solution 2-fluorobenzaldehyde(0.59 mL, 5.60 mmol) was added and the reaction allowed to stand at roomtemperature for 48 h. The forming yellow crystals were filtered off,washed with EtOH abs and dried under vacuum. Further purification wasnot necessary.

[0185] Yellow crystals (91%). ¹H NMR (400 MHz, d₆-DMSO) δ 10.11 (1H,s-br), 7.90 (2H, s), 7.57 (2H, qd, J=7.6 Hz, J=1.6 Hz), 7.51 (2H, td,J=8.0 Hz, J=1.2 Hz), 7.37 (4H, q, J=10.0 Hz), 4.37 (4H, s), 3.60-3.20(1H, s-br), 1.91 (3H, s). ¹³C NMR (100 MHz, d₆-DMSO) δ 181.95, 172.04,160.33 (d, J=249 Hz), 132.57 (d, J=9 Hz), 131.81 (d, J=4 Hz), 131.05,129.86, 124.96 (d, J=3 Hz), 121.49 (d, J=13 Hz), 116.1 (d, J=21 Hz),43.79, 21.11. HREIMS: m/z 311.1123 (M⁺-HOAc, C₁₉H₁₅NOF₂ requires311.1122).

Example 32 Cell Viability and VEGF/TF Inhibition Analysis

[0186] As described above, the effect of the compounds of the presentinvention on Neutral Red uptake, VEGF production, and TF production wasmeasured for a variety of human cancer cell lines. Some of this data isalso summarized in FIGS. 1-5. Note that, as applied to compounds EF4,MD6 and MD10 in the figures, the term “known” means that the listedcompounds have appeared in the literature, but without any suggestionthat those compounds exhibit anti-angiogenic properties or usefulness asa cancer treatment. Table 1 below lists the results for selectedcurcumin analogs of the present invention in comparison to the resultsfor curcumin and other known chemotherapeutic and anti-angiogenic agentsfor RPMI-7951 cells. TABLE 1 Characteristics of Selected Novel CurcuminAnalogs in the Human Melanoma Cell Line RPMI-7951 as Measured by theNeutral Red, VEGE ELISA^(a) and TF ELISA^(d) Assays; Comparison withMelanoma and Anti-Angiogenesis Agents Neutral Red VEGF^(c) TF^(C)Uptake,%^(b) ELISA ELISA Compound 5 μM 20 μM 5 μM 20 μM 5 μM 20 μM DMSO(0.1%) 100 3713 7054 Curcumin 100 14 3989 714 7974 1026 Series I MD6 976 2826 273 1753 ND MD1O 82 6 1585 315 1793 ND EF-1 92 8 1923 234 ND NDEF-2 46 7 1295 84 1009 ND EF-3 77 8 2092 230 ND ND EF-4 98 4 2159 199 NDND EF-8 97 8 2208 319 ND ND EF-9 88 11 1868 705 ND ND EF-10 ND 8 ND 257ND ND Series II MD279L 100 100 2235 1603 5791 5858 MD279U 100 100 2511779 7200 5316 EF-15 100 100 2361 894 7506 7663 Melanoma ChemotherapeuticAgents Decabazine 100 100 1136 1531 ND ND Cisplatin 100 100 KnownAnti-angiogenic Agent Thalidomide 100 100 1810 2827 ND ND

[0187] As indicated in the table and accompanying figures, two series ofanalogs were discovered. The Series I analogs proved to inhibit VEGFproduction and simultaneously inhibit cell growth for several cancercell lines. Some of the compounds within this group were also moreefficacious than TAXOL in preventing growth of a human breast cancercell line and more potent than curcumin and CISPLATIN in inhibitingproliferation of normal human and transformed murine VECs.

[0188] The Series II analogs, which include EF15, EF19-22, MD279L andMD279U, selectively blocked VEGF production without causing cell death.These compounds were also not cytotoxic to normal or malignant VECs.

[0189] These results indicate that the analogs of the present inventioncan directly inhibit tumor and vascular endothelial cell growth as wellas shut down the production of VEGF which is vital for tumor-inducedangiogenesis. Thus, the results suggest that the novel Series I analogsare potential anti-cancer/anti-angiogenic agents, while the Series IIcompounds are promising anti-angiogenic drugs with little toxicity tonormal VECs.

Example 33

[0190] TABLE 2 Vascular Endothelial Growth Factor (VEGF) Production byHuman Melanoma Cell Line, RPMI-795 1 measured by VEGF ELISA assay CellViability VEGF Concentrations 5 μM 20 μM 5 μM 20 μM of compounds (%) (%)(pg/ml) (%) (pg/ml) (%) DM50 (0.1%) 100 100 4197 100  4197 100 Curcumin105 47 4111 98 221 5 Our synthetic curcumin analogs which inhibit VEGFproduction and cell growth. EF-1 98 43 2573 61 0 0 EF-2 68 42 1522 36 00 EF-3 88 43 2827 67 0 0 EF-4 102 38 2920 70 0 0 EF-5 n.d. 84 n.d. n.d.n.d. 41 EF-6 n.d. 72 n.d. n.d. n.d. 45 EF-7 n.d. 78 n.d. n.d. n.d. 27EF-8 94 54 2276 54 35 1 EF-9 92 70 1925 46 595 14 EF-10 n.d. 54 n.d.n.d. 217 5 EF-11 n.d. 93 n.d. n.d. n.d. 51 EF-12 n.d. 92 n.d. n.d. n.d.70 EF-13 n.d. 92 n.d. n.d. n.d. 53 EF-14 n.d. 86 mcI. n.d. n.d. 36 EF-2512 n.d.  216* 21 n.d. n.d. A231L n.d. 77 n.d. n.d. n.d. 25 A231U n.d. 80n.d. n.d. n.d. 34 A232 n.d. 78 n.d. n.d. n.d. 40 A239 n.d. 86 n.d. n.d.n.d. 40 Our synthetic curcumin analogs which inhibit VEGF production,but not cell growth. EF-15 112 111 1187 28 2938 22 A279L 110 109 904 220 0 A279U 112 113 1197 28 1179 18 Drugs Currently in Market**Decarbazine 91 99 1171 28 1293 31 ***Thalidomide 105 97 1865 44 238757 Other Tested Compounds MD6 101 42 3200  76 30 1 MD10 91 42 1866  44 00 BA3 93 102 4860  116 2035 48 BA4 90 84 4708  112 3348 80

Example 34

[0191] TABLE 3 Vascular Endothelial Growth Factor (VEGF) and TissueFactor (TF) Production by Human Prostate Cancer Cell lines, DU-145 andPC-3 measured by VEGF ELISA and TF ELISA, respectively All compounds areused at 20 μM and DM50 (solvent control), 0.1% at a final concentration.VEGF TF DU-145 PC-3 DU-145 PC-3 pg/ml pg/ml pg/ml pg/ml DMSO 18760 ±2633  2103 ± 100  7699 ± 406  203 ± 6  (100%) (100%) (100%) (100%)Curcumin 17957 ± 1910  2088 ± 103  7134 ± 301  138 ± 10  (96%) (99%)(93%) (68%) Our synthetic curcumin analogs A 26565 ± 9818  1729 ± 59 n.d. n.d. (142%) (82%) B 15321 ± 5607  2168 ± 279  n.d. n.d. (82%) (99%)C 20559 ± 9224  1734 ± 397  n.d. n.d. (110%) (82%) EF-1 19616 ± 4624 325 ± 75  12879 ± 107 ± 9  (105%) (15%) 149 (167%) (53%) EF-2 7516 ±1915 26 ± 44 5540 ± 364  73 ± 10 (40%) (1%) (72%) (36%) EF-3 25901 ±620  917 ± 261 6687 ± 188  140 ± 8  (138%) (44%) (87%) (69%) EF-4 10274± 4467  48 ± 83 5180 ± 420  33 ± 7  (55%) (2%) (67%) (16%) A231U 27875 ±4446  984 ± 349 n.d. n.d. (149%) (47%) A271a 24190 ± 2160  1196 ± 438n.d. n.d. (129%) (57%) MD6 18428 ± 3377  1820 ± 283  n.d. n.d. (98%)(87%) MD10 7830 ± 2262 611 ± 248 5886 ± 332  0 ± 0 (42%) (29%) (76%)(0%) BA-8 40883 ± 6639  1648 ± 229  n.d. n.d. (218%) (78%)

Example 35

[0192] TABLE 4 Vascular Endothelial Growth Factor (VEGF) Production byHuman Prostate Cancer Cell Lines, DU-145 & PC-3 measured by VEGF ELISAassay All compounds are used at 20 μM and DMSO (solvent control), 0.1%at a final concentration. DU-145 PC-3 VEGF VEGF (pg/ml) (%) (pg/ml) (%)DMSO 18760 ± 2633 100 2103 ± 100  100 Curcumin 17957 ± 1910 96 2088 ±103  99 Our synthetic curcumin analogs A 26565 ± 9818 142 1729 ± 59  82B 15321 ± 5607 82 2168 ± 279  103 C 20559 ± 9224 110 1734 ± 397  82 EF-119616 ± 4624 105 325 ± 75  15 EF-2  7516 ± 1915 40 26 ± 44 1 EF-3 25901± 620  149 917 ± 261 44 EF-4 10274 ± 4467 55 48 ± 83 2 EF-5 20140 ± 2874107 2748 ± 416  131 A231L 27927 ± 4466 149 1687 ± 250  80 A231U 27875 ±4446 149 984 ± 349 47 A232 25308 ± 4722 135 1955 ± 437  93 A239 27229 ±2148 145 1678 ± 293  80 A271a 24190 ± 2160 129 1196 ± 438  57 A272 24155± 4635 129 1864 ± 339  89 A277a 27403 ± 3143 146 2007 ± 30  95 A277b27124 ± 1346 145 1868 ± 38  89 MD6 18428 ± 3377 98 1820 ± 283  87 MD10 7830 ± 2262 42 611 ± 248 29 BA-S 40883 ± 6639 218 1648 ± 229  78

Example 36

[0193] TABLE 5 Tissue Factor (TF) Production by Human Melanoma CellLine, RPMJ-7951 measured by TF ELISA assay TF Concentrations 1 μM 5 μM20 μM of compounds (pg/ml) (%) (pg/ml) (%) (pg/ml) (%) DM50 (0.1%) 7054100 7054 100 7054 100 Curcumin n.d. 8871 126 1026 15 Our syntheticcurcumin analogs EF-2 n.d. 5780 82 1009 14 EF-15 n.d. 7506 106 7663 109EF-32 n.d. n.d. 8654 123 A231U n.d. 5473 78 5981 85 A279L n.d. 5791 825858 83 A279U n.d. 7200 102 5316 75 MD6 6400 91 1753 25 n.d. MD1O 7966113 1793 25 n.d. BA-i n.d. n.d. 6669 95 BA-3 n.d. n.d. 2088 30 BA-4 n.d.7043 100 2120 30

Example 37

[0194] TABLE 6 Tissue Factor (TF) Production by Human Breast Cancer CellLine, MDA-MB-231 measured by TF ELISA assay TF Concentrations 0.5 μM 5μM 10 μM 20 μM of compounds (pg/ml) (%) (pg/ml) (%) (pg/ml) (%) (pg/ml)(%) DMSO (0.1%) 45753 100 45753 100 45753 100 45753 100 Curcumin n.d.45814 100 44727 98 n.d. Our synthetic curcumin analogs EF-2 37697 82n.d. n.d. n.d. A279L n.d. n.d. n.d. 42601 93 A279L n.d. n.d. n.d. 4219092 EF-15 n.d. n.d. n.d. 46394 101

Example 38

[0195] A number of the compounds of the present invention were screenedfor activity using the NCI Anti-Tumor Screen. The results are given inTables 7-9 below. Sixty human tumor cell lines were treated for 48 hourswith 10-fold dilutions of compounds at a minimum of five concentrations(0.01 μM-100 μM). Sulforhodamine B (SRB) assay was used to calculatecell viability or growth. GI50 refers to the concentration at which thedrug inhibits tumor cell growth by 50%. LC50 refers to the concentrationat which the drug causes 50% tumor cell death. EF 24 and EF 25 representthe average of three separate experiments. TABLE 7 Median GrowthInhibitory Concentration (GI50, μM) of Compounds in NCI Anti-TumorScreen COMPOUNDS Panel/Cell Line EF4 EF7 EF9 EF11 EF15 EF19 EF24 EF25Curcumin CISPLATIN Leukemia CCRF-CEM 1.4 12.5 3.1 22.0 41.5 18.2 0.2 0.33.2 0.2 HL-60 (TB) 1.9 17.7 4.2 21.3 49.9 18.1 0.4 1.1 7.9 0.1 K-562 2.514.9 3.5 27.4 78.5 19.0 0.3 0.6 3.2 1.0 MOLT-4 2.2 7.7 9.3 19.7 >100 2.60.5 0.4 5.0 0.3 RPMI-8226 0.6 14.8 2.4 18.2 49.8 21.7 0.2 0.2 2.0 0.5 SR1.6 15.2 1.2 21.5 52.0 22.7 0.1 0.2 3.2 0.1 Non-Small Cell Lung CancerA549 2.6 14.9 8.9 38.7 >100 18.0 1.9 2.5 12.6 0.8 EKVX 2.3 17.3 11.830.6 64.0 17.5 0.9 1.4 15.8 1.6 HOP-62 2.9 16.3 16.6 83.3 >100 18.0 0.71.5 4.0 0.3 HOP-92 2.8 12.7 16.3 23.5 >100 17.5 3.0 2.4 ND 0.6 NCI-H22619.0 18.7 24.3 >100 >100 19.9 3.6 2.2 20.0 0.8 NCI-H23 1.8 17.6 12.335.8 >100 22.8 0.9 1.7 5.0 0.1 NCI-H322M 2.7 16.9 16.7 36.5 >100 20.31.4 1.9 15.8 1.3 NCI-H460 2.3 17.6 12.7 30.7 >100 17.9 81.0 1.2 7.9 0.1NCI-H522 2.9 16.2 13.4 33.3 69.9 18.2 0.9 0.9 5.0 0.8 Colon Cancer COLO205 2.0 17.5 7.6 39.6 >100 17.5 0.8 2.0 12.6 4.0 HCC-2998 1.7 18.4 3.320.5 31.6 17.7 1.1 1.5 3.2 0.1 HCT-116 2.2 17.6 1.9 36.8 >100 17.8 0.20.8 3.2 1.3 HCT-15 2.2 17.2 4.9 42.6 49.1 24.8 0.2 0.7 4.0 1.6 HT29 2.217.8 4.1 45.9 46.1 17.9 0.3 2.0 5.0 1.3 KM12 1.6 17.1 2.3 19.1 55.7 17.60.2 0.3 5.0 2.0 SW-620 2.7 16.2 3.2 32.5 79.8 17.7 0.2 0.5 4.0 0.8 CNSCancer SF-268 3.0 19.8 13.7 24.6 >100 21.5 0.6 1.4 6.3 0.2 SF-295 3.415.5 14.2 53.8 80.9 18.7 85.0 1.7 7.9 0.3 SF-539 2.0 17.3 2.6 39.9 >10016.9 0.3 0.5 3.2 0.4 SNB-19 3.2 17.5 14.7 48.0 >100 16.1 0.7 2.2 7.9 1.0SNB-75 2.0 11.8 11.2 20.3 98.8 19.0 2.0 1.8 6.3 0.6 U251 2.0 16.1 2.135.7 95.1 17.4 0.2 0.7 5.0 0.4 Melanoma LOX IMVI 2.2 17.2 3.4 40.3 >10020.0 0.2 0.2 2.5 0.3 MALME-3M 2.3 17.5 17.8 40.7 >100 18.0 2.3 1.9 12.60.2 M14 1.9 15.7 4.1 33.9 >100 18.3 1.2 1.8 4.0 0.3 SK-MEL-2 2.4 16.814.5 24.0 31.7 18.4 2.6 2.0 15.8 1.3 SK-MEL-28 2.9 17.4 18.6 42.5 83.717.1 2.1 2.4 5.0 0.8 SK-MEL-5 1.6 15.5 12.1 23.2 53.9 17.1 1.3 1.5 7.90.5 UACC-257 2.9 16.9 16.0 29.8 55.4 17.9 1.8 1.6 12.6 1.0 UACC-62 1.512.2 13.9 21.3 66.7 15.1 1.6 1.7 6.3 0.3 Ovarian Cancer IGROV1 2.9 15.93.8 35.9 >100 17.2 0.8 0.8 7.9 0.6 OVCAR-3 3.7 15.6 14.1 41.8 >100 17.91.1 0.9 6.3 1.0 OVCAR-4 2.5 14.1 15.2 28.7 >100 17.4 0.7 2.0 10.0 0.3OVCAR-5 2.6 16.7 20.0 89.2 >100 18.5 1.3 1.7 15.8 1.3 OVCAR-8 2.9 17.03.4 47.1 >100 18.0 0.4 0.8 7.9 1.0 SK-OV-3 6.7 16.6 15.4 >100 >100 18.51.7 2.4 7.9 1.3 Renal Cancer 786-0 2.3 16.4 3.5 41.3 >100 17.0 0.2 0.43.2 0.3 A498 2.2 17.2 2.4 34.2 23.3 16.3 0.9 1.3 15.8 2.0 ACHN 3.3 15.716.5 41.0 >100 17.8 1.1 1.5 12.6 0.5 CAKI-1 3.1 14.3 15.0 29.2 >100 21.80.8 1.8 12.6 0.3 RXF 393 0.5 11.0 2.3 39.0 23.5 18.8 0.6 0.6 3.2 0.8SN12C 2.3 15.2 17.1 34.2 >100 17.4 1.0 1.4 7.9 1.0 TK-10 3.0 17.4 17.941.2 >100 17.4 3.1 2.2 15.8 1.3 UO-31 1.6 13.5 9.6 28.4 96.8 18.2 0.61.2 12.6 0.8 Prostate Cancer PC-3 2.1 17.2 4.8 34.9 >100 17.5 0.3 0.57.9 1.6 DU-145 1.4 16.4 2.1 29.9 >100 18.1 0.7 1.2 15.8 0.4 BreastCancer MCF-7 2.7 18.0 3.0 28.7 44.6 17.7 0.2 0.3 3.2 0.4 NCI/ADR-RES 2.719.3 30.7 86.1 >100 25.3 0.9 1.6 7.9 0.8 MDA-MB-231 3.5 19.8 14.125.6 >100 22.4 1.3 1.9 20.0 3.2 HS 578T 5.1 22.4 9.9 52.1 >100 21.9 1.05.1 10.0 1.3 MDA-MB-435 1.8 17.5 3.2 31.4 61.2 17.1 0.6 1.0 3.2 1.6MDA-N 1.6 16.1 2.9 36.9 67.8 16.9 0.5 1.0 2.5 0.8 BT-549 2.4 17.1 6.532.9 >100 17.9 0.7 1.5 5.0 1.3 T-47D 1.7 7.7 4.4 24.7 18.5 15.5 1.3 1.37.9 2.0 Mean 2.4 15.8 7.2 34.7 75.9 17.8 0.7 1.1 6.7 0.7

[0196] As shown, EF4, EF24 and EF 25 exhibited a lower GI50 than thechemotherapeutic agent CISPLATIN for several cell types. TABLE 8 MedianLethal Concentration (LC50, μM) of Compounds in NCI Anti-Tumor ScreenCOMPOUNDS Panel/Cell Line EF4 EF7 EF9 EF19 EF24 EF25 Curcumin CISPLATINLeukemia CCRF-CEM 59.1 79.5 >100 72.3 70.9 >100 <100 39.8 HL-60 (TB)53.5 76.0 >100 66.8 >100 >100 <100 50.1^(#) K-562 91.4 70.7 >100 90.175.4 91.8 <100 50.1 MOLT-4 69.5 64.7 >100 75.6 66.2 >100 79.4 50.1RPMI-8226 >100 92.7 95.6 >100 54.9 70.9 <100 50.1 SR >100 76.8 77.9 82.668.2 76.3 79.4 50.1 Non-Small Cell Lung Cancer A549 >100 53.3 71.0 65.889.4 71.8 79.4 50.1 EKVX 37.2 56.6 49.1 57.7 9.4 24.2 79.4 50.1 HOP-6269.1 63.1 60.7 58.7 28.8 37.2 63.1 50.1 HOP-92 41.4 54.9 70.4 59.3 23.335.4 ND 50.1 NCI-H226 >100 78.8 >100 75.7 48.3 58.0 <100 50.1 NCI-H239.4 62.9 62.3 96.8 5.3 12.9 <100 50.1 NCI-H322M 42.3 56.0 55.6 59.2 17.032.2 63.1 50.1 NCI-H460 40.2 66.8 60.9 57.5 50.1 38.9 63.1 39.8NCI-H522 >100 >100 57.1 75.4 22.9 14.1 79.4 50.1 Colon Cancer COLO 2058.7 55.9 52.8 55.9 8.0 33.4 63.1 50.1 HCT-2998 5.9 56.8 32.2 56.1 5.05.3 31.6 19.9 HCT-116 13.2 57.6 7.7 56.3 1.8 11.1 50.1 50.1 HCT-15 >10068.9 64.7 >100 13.6 16.1 79.4 50.1 HT29 29.1 60.6 41.9 62.6 57.3 67.5<100 50.1 KM12 6.5 55.5 14.6 56.1 3.4 10.7 63.1 50.1 SW-620 76.3 65.962.6 63.7 4.4 75.3 79.4 50.1 CNS Cancer SF-268 53.2 >100 >100 85.3 55.643.3 <100 50.1 SF-295 79.5 55.1 58.8 69.7 20.6 35.0 50.1 50.1 SF-53924.7 55.7 23.1 55.3 7.6 16.8 31.6 50.1 SNIB-19 42.3 55.9 58.8 57.3 19.337.3 63.1 50.1 SNB-75 21.5 49.0 48.8 57.5 8.8 14.0 50.1 50.1 U251 18.554.4 12.9 55.8 0.8 5.8 50.1 39.8 Melanoma LOX IMVI >100 73.4 67.6 74.46.7 3.6 <100 50.1 MALME-3M 24.3 68.8 73.4 59.6 25.2 19.3 50.1 39.8 M1413.2 54.0 48.6 68.7 27.7 19.4 50.1 50.1 SK-MEL-2 25.3 57.0 58.9 57.019.5 12.6 79.4 50.1 SK-MEL-28 33.8 62.9 61.1 55.6 33.1 6.6 50.1 50.1SK-MEL-5 5.7 53.7 53.1 55.5 6.4 6.6 50.1 7.9 UACC-257 57.8 55.2 62.156.3 13.5 8.2 50.1 50.1 UACC-62 6.1 49.6 52.8 53.2 4.5 14.4 50.1 15.8Ovarian Cancer IGROV1 43.9 60.9 46.2 55.6 7.1 14.5 79.4 50.1 OVCAR-340.7 54.5 52.1 59.3 16.2 23.5 63.1 50.1 OVCAR-4 18.4 53.1 61.6 65.1 17.223.3 <100 39.8 OVCAR-5 55.9 58.4 63.3 61.1 9.2 8.4 79.4 50.1OVCAR-8 >100 86.4 85.3 67.3 31.3 23.0 79.4 50.1 SK-OV-3 63.6 56.2 53.765.9 12.8 26.9 50.1 50.1 Renal Cancer 786-O 15.9 54.8 49.6 55.4 1.6 32.639.8 50.1 A498 16.2 55.6 23.0 54.7 3.4 6.0 63.1 50.1 ACHN 32.7 54.0 54.856.3 5.6 18.2 63.1 50.1 CAKI-1 >100 52.3 93.4 67.8 27.2 53.7 50.1 50.1RXF 393 30.8 49.2 17.9 58.4 3.6 4.6 50.1 50.1 SN12C 21.5 54.4 62.3 55.95.4 12.0 63.1 50.1 TK-10 22.6 55.9 56.5 56.3 27.8 24.6 63.1 50.1 UO-316.2 51.3 49.5 56.6 4.5 68.1 50.1 25.1 Prostate Cancer PC-3 28.6 55.641.0 56.7 7.2 24.8 63.1 50.1 DU-145 7.0 54.8 29.0 56.6 5.4 20.1 63.150.1 Breast Cancer MCF-7 44.1 65.9 42.2 57.1 5.0 44.7 <100 50.1NCI/ADR-RES >100 >100 >100 >100 60.6 71.0 <100 50.1 MDA-MB-231 >100 74.657.7 66.3 9.1 17.2 <100 50.1 HS 578T >100 >100 >100 >100 81.4 72.3 <10050.1 MDA-MB-435 8.0 56.0 35.9 55.6 7.4 16.3 39.8 39.8 MDA-N 20.2 56.933.2 55.2 5.6 20.0 31.6 39.8 BT-549 26.1 56.8 47.3 64.7 4.7 13.6 39.850.1 T-47D 50.8 55.4 56.1 68.9 51.9 92.9 <100 50.1 Mean 33.9 61.7 52.564.6 13.2 24.2 66.0 46.0

[0197] EF4, EF9, EF24, and EF25 exhibited a lower LC50 than thechemotherapeutic agent CISPLATIN for several cell types.

[0198] Human tumor and endothelial cell lines were treated for 72 hourswith compounds at a minimum of four concentrations between 0.1 μM-40 μM.Neutral Red assay was used to calculate cell viability. The numbers inTable 9 are representative of at least three separate experiments. TABLE9 Median Growth Inhibitory Concentration (GI50, μM) of Compounds inEmory Laboratory Cell Screen COMPOUNDS Panel/Cell Line EF2 EF4 EF25 EF34MD6 MD10 MD283 MD286 MD287 Curcumin Melanoma RPMI 7951 0.8 3.6 0.6 ND1.9 2.2 1.4 1.0 0.7 6.3 Breast Cancer MDA-MB-231 1.5 ND 0.8 0.8 ND ND1.8 ND ND 11.6 MDA-MB-435 3.3 ND 1.8 ND ND ND 1.9 ND ND 16.3 HUVECS 1.7ND 1.5 ND 3.8 14.0 3.1 2.3 6.8 25 Mean 1.8 3.6 1.2 0.8 2.9 8.1 2.1 1.73.8 14.8

[0199] Many modifications and other embodiments of the invention willcome to mind to one skilled in the art to which this invention pertainshaving the benefit of the teachings presented in the foregoingdescriptions and the associated drawings. Therefore, it is to beunderstood that the invention is not to be limited to the specificembodiments disclosed and that modifications and other embodiments areintended to be included within the scope of the appended claims.Although specific terms are employed herein, they are used in a genericand descriptive sense only and not for purposes of limitation.

That which is claimed:
 1. A compound of the Formula (I)

wherein: Y is OH, halogen, or CF₃; Z is H, OH, OR₁, halogen, or CF₃; X₁and X₂ are independently C or N; and A is selected from the groupconsisting of:

wherein n is 1-8; X₃ is O, S, SO, SO₂, NH, or NR₁; Q is NH or NR₁; andV₁₋₄ are each independently OH, OR₂, or halogen; R₁ and R₂ areindependently H, alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocycle, substitutedheterocycle, acyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl ordialkylaminocarbonyl; the dashed lines indicate the presence of optionaldouble bonds; and L is the point of bonding of A to the compoundstructure, with the proviso that Z is not H when Y is OH, Cl or Br and Ais

and pharmaceutically acceptable salts thereof.
 2. A compound accordingto claim 1, wherein Y is flourine.
 3. A compound selected from the groupconsisting of: 1,5-Bis-(2,4-difluorophenyl)penta-1,4-diene-3 -one;3,5-Bis-(2-fluorobenzylidene)-piperidin-4-one-acetate; and3,5-Bis-(2-hydroxybenzylidene)tetrahydro-4-H-pyran-4-one.
 4. Apharmaceutical formulation comprising a compound of claim 1 in apharmaceutically acceptable carrier.
 5. A method of treating canceroustissue in a subject, comprising administering to the subject aneffective amount of a compound of formula (I)

wherein: Y is OH, halogen, or CF₃; Z is H, OH, OR₁, halogen, or CF₃; X₁and X₂ are independently C or N; and A is selected from the groupconsisting of:

wherein n is 1-8; X₃ is O, S, SO, SO₂, NH, or NR₁; Q is NH or NR₁; andV₁₋₄ are each independently OH, OR₂, or halogen; R₁ and R₂ areindependently H, alkyl, substituted alkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocycle, substitutedheterocycle, acyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl ordialkylaminocarbonyl; the dashed lines indicate the presence of optionaldouble bonds; and L is the point of bonding of A to the compoundstructure, with the proviso that Z is not H when Y is OH, Cl or Br and Ais

and pharmaceutically acceptable salts thereof.
 6. A method according toclaim 5, wherein the effective amount comprises an amount sufficient toinhibit VEGF production in the cancerous tissue.
 7. A method accordingto claim 5, wherein the effective amount comprises an amount sufficientto inhibit TF production in the cancerous tissue.
 8. A method accordingto claim 5, wherein said administering step comprises administering aneffective amount of the compound in a pharmaceutically acceptablecarrier.
 9. A method of treating cancerous tissue in a subject,comprising administering to the subject an effective amount of acompound of formula (II)

wherein: X₄ is (CH₂)_(m), O, S, SO, SO₂, or NR₁₂, where R₁₂ is H, alkyl,substituted alkyl, acyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl or dialkylaminocarbonyl; m is 1-7; each X₅ isindependently N or C—R₁₁; and R₃-R₁₁ are independently H, halogen,hydroxyl, alkoxy, CF₃, alkyl, substituted alkyl, alkenyl, alkynyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, alkaryl,arylalkyl, heteroaryl, substituted heteroaryl, heterocycle, substitutedheterocycle, amino, alkylamino, dialkylamino, carboxylic acid,carboxylic ester, carboxamide, nitro, cyano, azide, alkylcarbonyl, acyl,or trialkylammonium; and the dashed lines indicate optional doublebonds; with the proviso that when X₄ is (CH₂)_(m), m is 2-6, and each X₅is C—R₁₁, R₃-R₁₁ are not alkoxy, and when X₄ is NR₁₂ and each X₅ is N,R₃-R₁₀ are not alkoxy, alkyl, substituted alkyl, alkenyl, alkynyl,cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, alkaryl,arylalkyl, heteroaryl, substituted heteroaryl, amino, alkylamino,dialkylamino, carboxylic acid, or alkylcarbonyl.
 10. A method accordingto claim 9, wherein the effective amount comprises an amount sufficientto inhibit VEGF production in the cancerous tissue.
 11. A methodaccording to claim 9, wherein the effective amount comprises an amountsufficient to inhibit TF production in the cancerous tissue.
 12. Amethod according to claim 9, wherein said administering step comprisesadministering an effective amount of the compound in a pharmaceuticallyacceptable carrier.